Substituted bis(pyridin-2-yl)amine derivatives, compositions thereof and pharmaceutical uses

Abstract

There is provided a substituted bis(pyridin-2-yl)amine derivative represented by formula (I), its pharmaceutically acceptable salts, solvates, stereoisomers, prodrugs, pharmaceutical compositions and its medical use. The derivative has significant CDK9 selective inhibitory activity and is very valuable in practice.

Description

Technical Field

[0001] The present invention relates to the field of pharmaceutical technology, and particularly relates to a substituted bis(pyridin-2-yl)amine derivative, a pharmaceutically acceptable salt, solvate, stereoisomer, prodrug, pharmaceutical composition thereof, and its medical use. Background Art

[0002] The cyclin-dependent kinase (CDK) protein family consists of members that are key regulators of the cell division cycle involved in the regulation of gene transcription (transcriptional CDK) (cell cycle CDK) and members with other functions. CDK requires activation by binding to a cyclin regulatory subunit. The cell cycle CDKs CDK1 / cyclin B, CDK2 / cyclin A, CDK2 / cyclin E, CDK4 / cyclin D, and CDK6 / cyclin D are successively activated to drive cells into and through the cell division cycle. The transcriptional CDKs CDK9 / cyclin T and CDK7 / cyclin H regulate the activity of RNA polymerase II by phosphorylation of the carboxyl-terminal domain (CTD).

[0003] CDK9 is the catalytic subunit in the positive transcription elongation factor b (P-TEFb) complex, regulates the transcriptional elongation of genes by phosphorylating the carbon-terminal region of RNA polymerase II, and is a kinase that regulates RNA transcriptional elongation located at chromosome 9q34.1. The CDK9 kinase is highly expressed in cardiomyocytes, hepatocytes, hematopoietic tissues, adipocytes, neurons, and muscle cells, and is generally highly expressed in tumor cells. CDK9 is also an important factor in tumor cell progression and maintenance. CDK9 is widely expressed in a variety of eukaryotic cells and human tissues. CDK9 inhibitors promote apoptosis of cancer cells by inhibiting the transcriptional elongation of genes to down-regulate the expression of related oncoproteins (MYC) and the expression of the apoptosis inhibitor protein Mcl-1. CDK9 inhibitors reactivate silenced genes by regulating the epigenetic factor BRG1, including the activation of ERVs in tumor cells, promote interferon expression, and make tumor cells more sensitive to immunotherapy.

[0004] Currently, several companies are developing CDK9 inhibitors, including the selective CDK9 inhibitor BAY1251152 developed by Bayer, the selective CDK9 inhibitor AZD4573 developed by AstraZeneca, the non-selective CDK9 inhibitor TP-1287 developed by Tolero, and the non-selective CDK9 inhibitor QHRD107 developed by Changzhou Qianhong Pharmaceutical Co., Ltd. However, most of the current selective CDK9 inhibitors are still in the early stage of clinical development. Among these drugs in clinical research, BAY1251152 has high toxicity and is administered by injection. The kinase selectivity and metabolism of AZD4573 are not good, restricting its better efficacy. Therefore, it is of important clinical significance to develop new highly active and highly selective CDK9 inhibitors that meet the clinical medication needs and improve patient compliance. SUMMARY OF THE INVENTION

[0005] The object of the present invention is to provide a substituted bis(pyridin-2-yl)amine derivative with high activity, good selectivity, excellent pharmacokinetic parameters, low toxicity, and suitable for oral administration.

[0006] In the first aspect of the present invention, there is provided a compound represented by formula (I), or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, or a solvate thereof, or a prodrug thereof:

[0007]

[0008] In the formula, (R 01 ) n is that the hydrogen on the pyridine ring is substituted by n R 01 , n is 0, 1, 2 or 3; each R 01 is the same or different and is independently selected from cyano, hydroxyl, carboxyl, halogen (preferably fluorine or chlorine), -C(O)OC 1-8 alkyl (preferably -C(O)OC 1-6 alkyl, more preferably -C(O)OC 1-3 alkyl), -OC(O)C 1-8 alkyl (preferably -OC(O)C 1-6 alkyl, more preferably -OC(O)C 1-3 alkyl), a 5- to 10-membered heteroaryl (preferably a 5- to 6-membered heteroaryl and an 8- to 10-membered heteroaryl), C 1-8 alkyl (preferably C 1-6 alkyl, more preferably C 1-3 alkyl) or C 1-8 alkoxy (preferably C 1-6 alkoxy, more preferably C 1-3 alkoxy); or two R 01Linking to form a 3- to 7-membered saturated or partially unsaturated monocyclic heterocycle or a 3- to 7-membered saturated or partially unsaturated monocyclic ring; wherein said C 1-8 alkyl, C 1-8 alkoxy, 3- to 7-membered saturated or partially unsaturated monocyclic heterocycle and 3- to 7-membered saturated or partially unsaturated monocyclic ring are unsubstituted or substituted with 1, 2 or 3 substituents each independently selected from the group consisting of: deuterium, halogen, cyano, hydroxy, carboxy, C 1-3 alkyl, C 1-3 alkoxy, C 2-4 alkenyl, C 2-4 alkynyl, halo C 1-3 alkyl, halo C 1-3 alkoxy, NR a0 R b0 、-SO 2 C 1-3 alkyl, -S(O)C 1-3 alkyl, -C(O)NR a0 R b0 、-C(O)OC 1-3 alkyl, -OC(O)C 1-3 alkyl, C 3-6 cycloalkyl, C 3-6 cycloalkoxy, 3- to 6-membered heterocycloalkyl, phenyl, 5- to 6-membered heteroaryl;

[0009] (R 02 ) m is such that the hydrogen on the pyridine ring is substituted by m R 02 groups, where m is 0, 1, 2 or 3; each R 02 is the same or different and is independently cyano, hydroxy, carboxy, halogen (preferably fluorine or chlorine), -C(O)OC 1-8 alkyl (preferably -C(O)OC 1-6 alkyl, more preferably -C(O)OC 1-3 alkyl), -OC(O)C 1-8 alkyl (preferably -OC(O)C 1-6 alkyl, more preferably -OC(O)C 1-3 alkyl), 5- to 10-membered heteroaryl (preferably 5- to 6-membered heteroaryl and 8- to 10-membered heteroaryl), C 1-8 alkyl (preferably C 1-6 alkyl, more preferably C 1-3 alkyl) or C 1-8 alkoxy (preferably C 1-6 alkoxy, more preferably C 1-3 alkoxy); or two R 02 groups link to form a 3- to 7-membered saturated or partially unsaturated monocyclic heterocycle or a 3- to 7-membered saturated or partially unsaturated monocyclic ring; wherein said C 1-8 alkyl, C1-8 The alkoxy group, 3- to 7-membered saturated or partially unsaturated monocyclic heterocycle, and 3- to 7-membered saturated or partially unsaturated monocyclic ring are unsubstituted or substituted with 1, 2, or 3 substituents each independently selected from the group consisting of deuterium, halogen, cyano, hydroxy, carboxy, C 1-3 alkyl, C 1-3 alkoxy, C 2-4 alkenyl, C 2-4 alkynyl, halo-C 1-3 alkyl, halo-C 1-3 alkoxy, NR a0 R b0 、-SO 2 C 1-3 alkyl, -S(O)C 1-3 alkyl, -C(O)NR a0 R b0 、-C(O)OC 1-3 alkyl, -OC(O)C 1-3 alkyl, C 3-6 cycloalkyl, C 3-6 cycloalkyloxy, 3- to 6-membered heterocycloalkyl, phenyl, 5- to 6-membered heteroaryl;

[0010] Q is a 5- to 10-membered heteroaryl, and the 5- to 10-membered heteroaryl is unsubstituted or substituted with 1, 2, or 3 substituents each independently selected from the group consisting of deuterium, halogen, cyano, hydroxy, carboxy, C 1-3 alkyl, C 1-3 alkoxy, C 2-4 alkenyl, C 2-4 alkynyl, halo-C 1-3 alkyl, halo-C 1-3 alkoxy, NR a0 R b0 、-SO 2 C 1-3 alkyl, -S(O)C 1-3 alkyl, -C(O)NR a0 R b0 、-C(O)OC 1-3 alkyl, -OC(O)C 1-3 alkyl, C 3-6 cycloalkyl, C 3-6 cycloalkyloxy, 3- to 6-membered heterocycloalkyl, substituted or unsubstituted phenyl, substituted or unsubstituted benzyl, substituted or unsubstituted 5- to 6-membered heteroaryl, substituted or unsubstituted -C 1-4 alkyl-5- to 6-membered heteroaryl; wherein substitution means that 1, 2, or 3 hydrogens in the group are independently substituted with substituents selected from the group consisting of halogen, cyano, hydroxy, carboxy, C 1-3 alkyl, C 1-3 alkoxy, hydroxy-substituted C 1-3Alkyl, NR a0 R b0 ;

[0011] Or Q is a structure represented by formula (a), formula (b) or formula (c):

[0012]

[0013] Wherein Z 1 Is CR a R b Or C(O); R a 、R b Are each independently hydrogen, cyano, hydroxy, carboxy, halogen (preferably fluorine or chlorine), -C(O)OC 1-8 Alkyl (preferably -C(O)OC 1-6 Alkyl, more preferably -C(O)OC 1-3 Alkyl), -OC(O)C 1-8 Alkyl (preferably -OC(O)C 1-6 Alkyl, more preferably -OC(O)C 1-3 Alkyl), 5- to 10-membered heteroaryl (preferably 5- to 6-membered heteroaryl and 8- to 10-membered heteroaryl), C 1-8 Alkyl (preferably C 1-6 Alkyl, more preferably C 1-3 Alkyl) or C 1-8 Alkoxy (preferably C 1-6 Alkoxy, more preferably C 1-3 Alkoxy); Or R a 、R b Together with the connected carbon atom form a 3- to 7-membered saturated or partially unsaturated monocyclic hetero ring or a 3- to 7-membered saturated or partially unsaturated monocyclic ring; Wherein the C 1-8 Alkyl, C 1-8 Alkoxy, 3- to 7-membered saturated or partially unsaturated monocyclic hetero ring and 3- to 7-membered saturated or partially unsaturated monocyclic ring are unsubstituted or substituted by 1, 2 or 3 substituents each independently selected from the group consisting of: deuterium, halogen, cyano, hydroxy, carboxy, C 1-3 Alkyl, C 1-3 Alkoxy, C 2-4 Alkenyl, C 2-4 Alkynyl, halo C 1-3 Alkyl, halo C 1-3 Alkoxy, NR a0 R b0 、-SO 2 C 1-3 Alkyl, -S(O)C 1-3 Alkyl, -C(O)NR a0 R b0 、-C(O)OC 1-3alkyl, -OC(O)C 1-3 alkyl, C 3-6 cycloalkyl, C 3-6 cycloalkyloxy, 3- to 6-membered heteroalkyl, phenyl, 5- to 6-membered heteroaryl;

[0014] Z 2 is (CR c R d ) t0 , -(CR c R d ) t1 -O- or -(CR c R d ) t2 -NR g -; each R c is the same or different, each R d is the same or different, and R c and R d are each independently hydrogen, cyano, hydroxy, carboxy, halogen (preferably fluorine or chlorine), -C(O)OC 1-8 alkyl (preferably -C(O)OC 1-6 alkyl, more preferably -C(O)OC 1-3 alkyl), -OC(O)C 1-8 alkyl (preferably -OC(O)C 1-6 alkyl, more preferably -OC(O)C 1-3 alkyl), 5- to 10-membered heteroaryl (preferably 5- to 6-membered heteroaryl and 8- to 10-membered heteroaryl), C 1-8 alkyl (preferably C 1-6 alkyl, more preferably C 1-3 alkyl) or C 1-8 alkoxy (preferably C 1-6 alkoxy, more preferably C 1-3 alkoxy); or R c and R d connected to the same carbon atom, or R c and R d connected to different carbon atoms, form a 3- to 7-membered saturated or partially unsaturated monocyclic heterocycle or a 3- to 7-membered saturated or partially unsaturated monocyclic ring; wherein the C 1-8 alkyl, C 1-8 alkoxy, 3- to 7-membered saturated or partially unsaturated monocyclic heterocycle and 3- to 7-membered saturated or partially unsaturated monocyclic ring are unsubstituted or substituted with 1, 2 or 3 substituents each independently selected from the group consisting of: deuterium, halogen, cyano, hydroxy, carboxy, C 1-3 alkyl, C 1-3 alkoxy, C 2-4 alkenyl, C 2-4 alkynyl, haloC1-3 alkyl, halo-C 1-3 alkoxy, NR a0 R b0 , -SO 2 C 1-3 alkyl, -S(O)C 1-3 alkyl, -C(O)NR a0 R b0 , -C(O)OC 1-3 alkyl, -OC(O)C 1-3 alkyl, C 3-6 cycloalkyl, C 3-6 cycloalkoxy, 3- to 6-membered heteroalkyl, phenyl, 5- to 6-membered heteroaryl;

[0015] R g is hydrogen or C 1-3 alkyl;

[0016] t0, t1, t2 are each independently 1, 2, 3, 4 or 5;

[0017] R 1a , R 2a are each independently hydrogen or C 1-3 alkyl; or R 1a is connected to R 2a to form a 5- to 8-membered saturated or partially unsaturated monocyclic heterocycle; the 5- to 8-membered saturated or partially unsaturated monocyclic heterocycle is unsubstituted or substituted by 1, 2, 3 or 4 R s1 ; wherein each R s1 is the same or different and is independently selected from: halogen, cyano, hydroxy, C 1-3 alkyl, C 1-3 alkoxy, halo-C 1-3 alkyl, halo-C 1-3 alkoxy; or any two R s1 on the same or different ring atoms are connected to form a 3- to 7-membered saturated or partially unsaturated monocyclic heterocycle or a 3- to 7-membered saturated or partially unsaturated monocyclic ring; the 3- to 7-membered saturated or partially unsaturated monocyclic heterocycle and the 3- to 7-membered saturated or partially unsaturated monocyclic ring are unsubstituted or substituted by 1, 2 or 3 substituents each independently selected from the group consisting of: deuterium, halogen, cyano, hydroxy, carboxy, C 1-3 alkyl, C 1-3 alkoxy, C 2-4 alkenyl, C 2-4 alkynyl, halo-C 1-3 alkyl, halo-C 1-3 alkoxy, NR a0 R b0 , -SO 2 C 1-3 alkyl, -S(O)C 1-3alkyl, -C(O)NR a0 R b0 、-C(O)OC 1-3 alkyl, -OC(O)C 1-3 alkyl, C 3-6 cycloalkyl, C 3-6 cycloalkyloxy, 3- to 6-membered heterocycloalkyl, phenyl, 5- to 6-membered heteroaryl;

[0018] R 3a 、R 4a each independently is hydrogen, cyano, C 1-8 alkyl (preferably C 1-6 alkyl, more preferably C 1-3 alkyl), -C(O)NR a0 R b0 or -C(O)OC 1-3 alkyl; or R 3a is connected to R 4a to form a 5- to 8-membered heterocycloalkenyl ring or a 5- to 6-membered heteroaryl ring; the 5- to 8-membered heterocycloalkenyl ring and the 5- to 6-membered heteroaryl ring are unsubstituted or substituted by 1, 2, 3 or 4 R s2 ; wherein each R s2 is the same or different and is independently selected from: halogen, cyano, hydroxy, C 1-3 alkyl, C 1-3 alkoxy, halo C 1-3 alkyl, halo C 1-3 alkoxy; or any two R s2 on the same ring atom or different ring atoms are connected to form a 3- to 7-membered saturated or partially unsaturated monocyclic heterocycle or a 3- to 7-membered saturated or partially unsaturated monocyclic ring; the 3- to 7-membered saturated or partially unsaturated monocyclic heterocycle and the 3- to 7-membered saturated or partially unsaturated monocyclic ring are unsubstituted or substituted by 1, 2 or 3 substituents each independently selected from the group consisting of: deuterium, halogen, cyano, hydroxy, carboxy, C 1-3 alkyl, C 1-3 alkoxy, C 2-4 alkenyl, C 2-4 alkynyl, halo C 1-3 alkyl, halo C 1-3 alkoxy, NR a0 R b0 、-SO 2 C 1-3 alkyl, -S(O)C 1-3 alkyl, -C(O)NR a0 R b0 、-C(O)OC 1-3 alkyl, -OC(O)C 1-3 alkyl, C 3-6 cycloalkyl, C 3-6Cycloalkyloxy, 3- to 6-membered heteroalkyl, phenyl, 5- to 6-membered heteroaryl;

[0019] R 5a is hydrogen or C 1-3 alkyl;

[0020] R 6a 、R 7a are each independently hydrogen or C 1-8 alkyl (preferably C 1-6 alkyl, more preferably C 1-3 alkyl); or R 6a is connected to R 7a to form a 5- to 8-membered heteroalkenyl ring or a 5- to 6-membered heteroaryl ring; the 5- to 8-membered heteroalkenyl ring and the 5- to 6-membered heteroaryl ring are unsubstituted or substituted with 1, 2, 3 or 4 R s3 ; wherein each R s3 is the same or different and is independently selected from: halogen, cyano, hydroxy, C 1-3 alkyl, C 1-3 alkoxy, halo C 1-3 alkyl, halo C 1-3 alkoxy; or any two R s3 on the same or different ring atoms are connected to form a 3- to 7-membered saturated or partially unsaturated monocyclic hetero ring or a 3- to 7-membered saturated or partially unsaturated monocyclic ring; the 3- to 7-membered saturated or partially unsaturated monocyclic hetero ring and the 3- to 7-membered saturated or partially unsaturated monocyclic ring are unsubstituted or substituted with 1, 2 or 3 substituents each independently selected from the group consisting of: deuterium, halogen, cyano, hydroxy, carboxy, C 1-3 alkyl, C 1-3 alkoxy, C 2-4 alkenyl, C 2-4 alkynyl, halo C 1-3 alkyl, halo C 1-3 alkoxy, NR a0 R b0 、-SO 2 C 1-3 alkyl, -S(O)C 1-3 alkyl, -C(O)NR a0 R b0 、-C(O)OC 1-3 alkyl, -OC(O)C 1-3 alkyl, C 3-6 cycloalkyl, C 3-6 cycloalkyloxy, 3- to 6-membered heteroalkyl, phenyl, 5- to 6-membered heteroaryl;

[0021] L is a bond or (CR e R f ) s ; each R eIdentical or different, each R f Identical or different, and R e and R f are each independently hydrogen, cyano, hydroxy, carboxy, halogen (preferably fluorine or chlorine), -C(O)OC 1-8 alkyl (preferably -C(O)OC 1-6 alkyl, more preferably -C(O)OC 1-3 alkyl), C 1-8 alkyl (preferably C 1-6 alkyl, more preferably C 1-3 alkyl) or C 1-8 alkoxy (preferably C 1-6 alkoxy, more preferably C 1-3 alkoxy); or R e , R f together with the attached carbon atom form a 3- to 7-membered saturated or partially unsaturated monocyclic heterocycle or a 3- to 7-membered saturated or partially unsaturated monocyclic ring; wherein the C 1-8 alkyl, C 1-8 alkoxy, 3- to 7-membered saturated or partially unsaturated monocyclic heterocycles and 3- to 7-membered saturated or partially unsaturated monocyclic rings are unsubstituted or substituted by 1, 2 or 3 substituents each independently selected from the group consisting of deuterium, halogen, cyano, hydroxy, carboxy, C 1-3 alkyl, C 1-3 alkoxy, C 2-4 alkenyl, C 2-4 alkynyl, halo-C 1-3 alkyl, halo-C 1-3 alkoxy, NR a0 R b0 , -SO 2 C 1-3 alkyl, -S(O)C 1-3 alkyl, -C(O)NR a0 R b0 , -C(O)OC 1-3 alkyl, -OC(O)C 1-3 alkyl, C 3-6 cycloalkyl, C 3-6 cycloalkyloxy, 3- to 6-membered heterocycloalkyl, phenyl, 5- to 6-membered heteroaryl;

[0022] s is 1, 2 or 3;

[0023] R 1 , R 2 , R 3 are each independently hydrogen, cyano, hydroxy, carboxy, halogen (preferably fluorine or chlorine), -C(O)OC 1-8 alkyl (preferably -C(O)OC 1-6 alkyl, more preferably -C(O)OC1-3 alkyl), C 1-8 alkyl (preferably C 1-6 alkyl, more preferably C 1-3 alkyl) or C 1-8 alkoxy (preferably C 1-6 alkoxy, more preferably C 1-3 alkoxy); wherein the C 1-8 alkyl, C 1-8 alkoxy is unsubstituted or substituted by 1, 2 or 3 substituents each independently selected from the group consisting of: deuterium, halogen, cyano, hydroxy, carboxy, C 1-3 alkyl, C 1-3 alkoxy, C 2-4 alkenyl, C 2-4 alkynyl, halo C 1-3 alkyl, halo C 1-3 alkoxy, NR a0 R b0 、-SO 2 C 1-3 alkyl, -S(O)C 1-3 alkyl, -C(O)NR a0 R b0 、-C(O)OC 1-3 alkyl, -OC(O)C 1-3 alkyl, C 3-6 cycloalkyl, C 3-6 cycloalkyloxy, 3- to 6-membered heteroalkyl, phenyl, 5- to 6-membered heteroaryl;

[0024] R 4 、R 5 are each independently hydrogen, cyano, hydroxy, carboxy, halogen (preferably fluorine or chlorine), -C(O)OC 1-8 alkyl (preferably -C(O)OC 1-6 alkyl, more preferably -C(O)OC 1-3 alkyl), C 1-8 alkyl (preferably C 1-6 alkyl, more preferably C 1-3 alkyl) or C 1-8 alkoxy (preferably C 1-6 alkoxy, more preferably C 1-3 alkoxy); wherein the C 1-8 alkyl, C 1-8 alkoxy is unsubstituted or substituted by 1, 2 or 3 substituents each independently selected from the group consisting of: deuterium, halogen, cyano, hydroxy, carboxy, C 1-3 alkyl, C 1-3 alkoxy, C 2-4 alkenyl, C 2-4 alkynyl, halo C 1-3Alkyl, halo-C 1-3 Alkoxy, NR a0 R b0 , -SO 2 C 1-3 Alkyl, -S(O)C 1-3 Alkyl, -C(O)NR a0 R b0 , -C(O)OC 1-3 Alkyl, -OC(O)C 1-3 Alkyl, C 3-6 Cycloalkyl, C 3-6 Cycloalkoxy, 3- to 6-membered heteroalkyl, phenyl, 5- to 6-membered heteroaryl;

[0025] Or R 4 Is linked to R 5 To form a fused 3- to 7-membered saturated or partially unsaturated monocyclic heterocycle or a fused 3- to 7-membered saturated or partially unsaturated monocyclic ring; the fused 3- to 7-membered saturated or partially unsaturated monocyclic heterocycle and the fused 3- to 7-membered saturated or partially unsaturated monocyclic ring are unsubstituted or substituted by 1, 2, 3 or 4 R s4 ; wherein each R s4 Is the same or different and is independently selected from: halogen, cyano, hydroxy, C 1-3 Alkyl, C 1-3 Alkoxy, halo-C 1-3 Alkyl, halo-C 1-3 Alkoxy; or any two R s4 On the same ring atom or different ring atoms are linked to form a 3- to 6-membered saturated or partially unsaturated monocyclic heterocycle or a 3- to 6-membered saturated or partially unsaturated monocyclic ring; the 3- to 6-membered saturated or partially unsaturated monocyclic heterocycle and the 3- to 6-membered saturated or partially unsaturated monocyclic ring are unsubstituted or substituted by 1, 2 or 3 substituents each independently selected from the group consisting of: deuterium, halogen, cyano, hydroxy, carboxy, C 1-3 Alkyl, C 1-3 Alkoxy, C 2-4 Alkenyl, C 2-4 Alkynyl, halo-C 1-3 Alkyl, halo-C 1-3 Alkoxy, NR a0 R b0 , -SO 2 C 1-3 Alkyl, -S(O)C 1-3 Alkyl, -C(O)NR a0 R b0 , -C(O)OC 1-3 Alkyl, -OC(O)C 1-3 Alkyl, C 3-6 Cycloalkyl, C 3-6Cycloalkyloxy, 3- to 6-membered heteroalkyl, phenyl, 5- to 6-membered heteroaryl; and

[0026] R a0 and R b0 are each independently hydrogen, C 1-3 alkyl or acetyl; or R a0 and R b0 together with the attached nitrogen atom form a 4- to 6-membered saturated monocyclic heterocycle; the 4- to 6-membered saturated monocyclic heterocycle is optionally substituted with 1, 2, or 3 substituents each independently selected from the group consisting of deuterium, halogen, cyano, hydroxy, carboxy, C 1-3 alkyl, C 1-3 alkoxy, C 2-4 alkenyl, C 2-4 alkynyl, halo C 1-3 alkyl, halo C 1-3 alkoxy, -SO 2 C 1-3 alkyl, -S(O)C 1-3 alkyl, -C(O)NR a0 R b0 -C(O)OC 1-3 alkyl, -OC(O)C 1-3 alkyl, C 3-6 cycloalkyl, C 3-6 cycloalkyloxy, 3- to 6-membered heteroalkyl.

[0027] In some embodiments, n is 1; R 01 is halogen (preferably fluorine).

[0028] In some embodiments, n is 1; R 01 is fluorine.

[0029] In some embodiments, is

[0030] In some embodiments, m is 0 or 1; R 02 is methoxy.

[0031] In some embodiments, m is 0.

[0032] In some embodiments, L is a bond or (CR e R f ) s ; R e and R f are each independently hydrogen.

[0033] In some embodiments, L is CH 2 .

[0034] In some embodiments, Z 1is CH 2 or C(O).

[0035] In some embodiments, Z 2 is (CH 2 ) t0 ; t0 is 1, 2, 3 or 4.

[0036] In some embodiments, formula (a) has the structure shown in formula (a1) or formula (a2):

[0037]

[0038] In some embodiments, formula (b) has the structure shown in formula (b1):

[0039]

[0040] wherein R 3a ', R 4a ' are each independently hydrogen, cyano, C 1-8 alkyl (preferably C 1-6 alkyl, more preferably C 1-3 alkyl), -C(O)NR a0 R b0 or -C(O)OC 1-3 alkyl; R 1a , R 2a , R a0 , R b0 are as defined in the specification.

[0041] In some embodiments, formula (b) has the structure shown in formula (b2):

[0042]

[0043] wherein E 1 is CR E1a R E1b , C(O) or SO 2 ;

[0044] E 2 is CR E2a R E2b , O, NR E2c , C(O) or SO 2 ;

[0045] E 3 is a bond, (CR E3a R E3b ) q , (CR E3a R E3b ) q -O, C(O) or SO 2 ;

[0046] R E1a 、R E1b Each independently is hydrogen, cyano, hydroxy, carboxy, halogen (preferably fluorine or chlorine), -C(O)OC 1-8 alkyl (preferably -C(O)OC 1-6 alkyl, more preferably -C(O)OC 1-3 alkyl), -OC(O)C 1-8 alkyl (preferably -OC(O)C 1-6 alkyl, more preferably -OC(O)C 1-3 alkyl), C 1-8 alkyl (preferably C 1-6 alkyl, more preferably C 1-3 alkyl) or C 1-8 alkoxy (preferably C 1-6 alkoxy, more preferably C 1-3 alkoxy); or R E1a 、R E1b together with the attached carbon atom form a 3- to 7-membered saturated or partially unsaturated monocyclic heterocycle or a 3- to 7-membered saturated or partially unsaturated monocyclic ring; wherein the C 1-8 alkyl, C 1-8 alkoxy, 3- to 7-membered saturated or partially unsaturated monocyclic heterocycle and 3- to 7-membered saturated or partially unsaturated monocyclic ring are unsubstituted or substituted with 1, 2 or 3 substituents each independently selected from the group consisting of deuterium, halogen, cyano, hydroxy, carboxy, C 1-3 alkyl, C 1-3 alkoxy, C 2-4 alkenyl, C 2-4 alkynyl, halo C 1-3 alkyl, halo C 1-3 alkoxy, NR a0 R b0 、-SO 2 C 1-3 alkyl, -S(O)C 1-3 alkyl, -C(O)NR a0 R b0 、-C(O)OC 1-3 alkyl, -OC(O)C 1-3 alkyl, C 3-6 cycloalkyl, C 3-6 cycloalkyloxy, 3- to 6-membered heterocycloalkyl, phenyl, 5- to 6-membered heteroaryl;

[0047] R E2a 、R E2b Each independently is hydrogen, cyano, hydroxy, carboxy, halogen (preferably fluorine or chlorine), -C(O)OC 1-8 alkyl (preferably -C(O)OC 1-6alkyl, more preferably -C(O)OC 1-3 alkyl), -OC(O)C 1-8 alkyl (preferably -OC(O)C 1-6 alkyl, more preferably -OC(O)C 1-3 alkyl), C 1-8 alkyl (preferably C 1-6 alkyl, more preferably C 1-3 alkyl) or C 1-8 alkoxy (preferably C 1-6 alkoxy, more preferably C 1-3 alkoxy); or R E2a , R E2b together with the attached carbon atom form a 3- to 7-membered saturated or partially unsaturated monocyclic heterocycle or a 3- to 7-membered saturated or partially unsaturated monocyclic ring; wherein the C 1-8 alkyl, C 1-8 alkoxy, 3- to 7-membered saturated or partially unsaturated monocyclic heterocycle and 3- to 7-membered saturated or partially unsaturated monocyclic ring are unsubstituted or substituted by 1, 2 or 3 substituents each independently selected from the group consisting of: deuterium, halogen, cyano, hydroxy, carboxy, C 1-3 alkyl, C 1-3 alkoxy, C 2-4 alkenyl, C 2-4 alkynyl, halo C 1-3 alkyl, halo C 1-3 alkoxy, NR a0 R b0 , -SO 2 C 1-3 alkyl, -S(O)C 1-3 alkyl, -C(O)NR a0 R b0 , -C(O)OC 1-3 alkyl, -OC(O)C 1-3 alkyl, C 3-6 cycloalkyl, C 3-6 cycloalkyloxy, 3- to 6-membered heterocycloalkyl, phenyl, 5- to 6-membered heteroaryl;

[0048] R E2c is hydrogen, C 1-8 alkyl (preferably C 1-6 alkyl, more preferably C 1-3 alkyl), C 3-8 cycloalkyl (preferably C 3-6 cycloalkyl), 3- to 8-membered heterocycloalkyl, -C 1-4 alkyl-3- to 8-membered heterocycloalkyl, -SO 2 C 1-8 alkyl (preferably -SO 2 C 1-6 alkyl, more preferably -SO2 C 1-3 alkyl), -C(O)NR a0 R b0 、-C(O)C 1-8 alkyl (preferably -C(O)C 1-6 alkyl, more preferably -C(O)C 1-3 alkyl), -C(O)OC 1-8 alkyl (preferably -C(O)OC 1-6 alkyl, more preferably -C(O)OC 1-3 alkyl) or C(O)NR a1 R b1 ; wherein said C 1-8 alkyl, C 3-8 cycloalkyl, 3- to 8-membered heterocycloalkyl, -SO 2 C 1-8 alkyl, -C(O)C 1-8 alkyl and -C(O)OC 1-8 alkyl are unsubstituted or substituted with 1, 2 or 3 substituents independently selected from the group consisting of: cyano, hydroxy, carboxy, halogen, acetyl, C 1-3 alkyl, C 1-3 alkoxy, C 2-4 alkenyl, C 2-4 alkynyl, -SO 2 C 1-3 alkyl, -S(O)C 1-3 alkyl, -C(O)NR a0 R b0 、-C(O)OC 1-3 alkyl, -OC(O)C 1-3 alkyl, -C(O)C 1-3 alkyl, -C(O)NR a1 R b1 、NR a1 R b1 、-NR c1 C(O)C 1-3 alkyl, -NR c1 C(O)C 3-6 cycloalkyl, 4- to 6-membered heterocycloalkyl;

[0049] Each R E3a is the same or different, each R E3b is the same or different, and R E3a and R E3b are each independently hydrogen, cyano, hydroxy, carboxy, halogen (preferably fluorine or chlorine), -C(O)OC 1-8 alkyl (preferably -C(O)OC 1-6 alkyl, more preferably -C(O)OC 1-3(alkyl), -OC(O)C 1-8 alkyl (preferably -OC(O)C 1-6 alkyl, more preferably -OC(O)C 1-3 alkyl), C 1-8 alkyl (preferably C 1-6 alkyl, more preferably C 1-3 alkyl) or C 1-8 alkoxy (preferably C 1-6 alkoxy, more preferably C 1-3 alkoxy); or R E3a and R E3b are linked to the same carbon atom, or R E3a and R E3b are linked to different carbon atoms to form a 3- to 7-membered saturated or partially unsaturated monocyclic heterocycle or a 3- to 7-membered saturated or partially unsaturated monocyclic ring; wherein the C 1-8 alkyl, C 1-8 alkoxy, 3- to 7-membered saturated or partially unsaturated monocyclic heterocycle and 3- to 7-membered saturated or partially unsaturated monocyclic ring are unsubstituted or substituted with 1, 2 or 3 substituents each independently selected from the group consisting of: deuterium, halogen, cyano, hydroxy, carboxy, C 1-3 alkyl, C 1-3 alkoxy, C 2-4 alkenyl, C 2-4 alkynyl, halo C 1-3 alkyl, halo C 1-3 alkoxy, -SO 2 C 1-3 alkyl, -S(O)C 1-3 alkyl, -C(O)NR a0 R b0 , -C(O)OC 1-3 alkyl, -OC(O)C 1-3 alkyl, C 3-6 cycloalkyl, C 3-6 cycloalkyloxy, 3- to 6-membered heterocycloalkyl, phenyl, 5- to 6-membered heteroaryl;

[0050] q is 1, 2 or 3;

[0051] R a1 , R b1 are each independently hydrogen, C 1-3 alkyl or C 3-6 cycloalkyl; R c1 is hydrogen or C 1-3 alkyl;

[0052] R 1a , R 2a , R a0 , R b0 as defined in the specification.

[0053] In some embodiments, formula (b2) has the structure shown in formula (b21):

[0054]

[0055] wherein R 1a ', R 2a ' are each independently hydrogen or C 1-3 alkyl; E 1 , E 2 , E 3 are as defined in formula (b2).

[0056] In some embodiments, formula (b2) has the structure shown in formula (b22):

[0057]

[0058] wherein W is (CR W1 R W2 ) u ; each R W1 is the same or different, each R W2 is the same or different, and R W1 and R W2 are each independently hydrogen, cyano, hydroxyl, carboxyl, halogen (preferably fluorine or chlorine), -C(O)OC 1-8 alkyl (preferably -C(O)OC 1-6 alkyl, more preferably -C(O)OC 1-3 alkyl), -OC(O)C 1-8 alkyl (preferably -OC(O)C 1-6 alkyl, more preferably -OC(O)C 1-3 alkyl), C 1-8 alkyl (preferably C 1-6 alkyl, more preferably C 1-3 alkyl) or C 1-8 alkoxy (preferably C 1-6 alkoxy, more preferably C 1-3 alkoxy); or R W1 and R W2 connected to the same carbon atom, or R W1 and R W2 connected to different carbon atoms form a 3- to 7-membered saturated or partially unsaturated monocyclic heterocycle or a 3- to 7-membered saturated or partially unsaturated monocyclic ring; wherein the C 1-8 alkyl, C 1-8An alkoxy group, a 3- to 7-membered saturated or partially unsaturated monocyclic heterocycle, and a 3- to 7-membered saturated or partially unsaturated monocyclic ring are unsubstituted or substituted by 1, 2, or 3 substituents each independently selected from the group consisting of: deuterium, halogen, cyano, hydroxy, carboxy, C 1-3 alkyl, C 1-3 alkoxy, C 2-4 alkenyl, C 2-4 alkynyl, halo-C 1-3 alkyl, halo-C 1-3 alkoxy, NR a0 R b0 , -SO 2 C 1-3 alkyl, -S(O)C 1-3 alkyl, -C(O)NR a0 R b0 , -C(O)OC 1-3 alkyl, -OC(O)C 1-3 alkyl, C 3-6 cycloalkyl, C 3-6 cycloalkoxy, 3- to 6-membered heterocycloalkyl, phenyl, 5- to 6-membered heteroaryl;

[0059] u is 1, 2, 3, or 4;

[0060] R a0 , R b0 as defined in the above specification;

[0061] E 1 , E 2 , E 3 as defined in formula (b2).

[0062] In some embodiments, formula (c) is the structure shown in formula (c1):

[0063]

[0064] wherein G 1 is CR G1a R G1b , C(O), or SO 2 ;

[0065] G 2 is CR G2a R G2b , O, NR G2c , C(O), or SO 2 ;

[0066] G 3 is a bond, (CR G3a R G3b ) p , C(O), or SO 2 ;

[0067] R G1a 、R G1b Each independently is hydrogen, cyano, hydroxy, carboxy, halogen (preferably fluorine or chlorine), -C(O)OC 1-8 alkyl (preferably -C(O)OC 1-6 alkyl, more preferably -C(O)OC 1-3 alkyl), -OC(O)C 1-8 alkyl (preferably -OC(O)C 1-6 alkyl, more preferably -OC(O)C 1-3 alkyl), C 1-8 alkyl (preferably C 1-6 alkyl, more preferably C 1-3 alkyl) or C 1-8 alkoxy (preferably C 1-6 alkoxy, more preferably C 1-3 alkoxy); or R G1a 、R G1b together with the attached carbon atom form a 3- to 7-membered saturated or partially unsaturated monocyclic heterocycle or a 3- to 7-membered saturated or partially unsaturated monocyclic ring; wherein the C 1-8 alkyl, C 1-8 alkoxy, 3- to 7-membered saturated or partially unsaturated monocyclic heterocycle and 3- to 7-membered saturated or partially unsaturated monocyclic ring are unsubstituted or substituted by 1, 2 or 3 substituents each independently selected from the group consisting of deuterium, halogen, cyano, hydroxy, carboxy, C 1-3 alkyl, C 1-3 alkoxy, C 2-4 alkenyl, C 2-4 alkynyl, halo C 1-3 alkyl, halo C 1-3 alkoxy, NR a0 R b0 、-SO 2 C 1-3 alkyl, -S(O)C 1-3 alkyl, -C(O)NR a0 R b0 、-C(O)OC 1-3 alkyl, -OC(O)C 1-3 alkyl, C 3-6 cycloalkyl, C 3-6 cycloalkyloxy, 3- to 6-membered heterocycloalkyl, phenyl, 5- to 6-membered heteroaryl;

[0068] R G2a 、R G2b Each independently is hydrogen, cyano, hydroxy, carboxy, halogen (preferably fluorine or chlorine), -C(O)OC 1-8 alkyl (preferably -C(O)OC 1-6Alkyl, more preferably -C(O)OC 1-3 alkyl), -OC(O)C 1-8 alkyl (preferably -OC(O)C 1-6 alkyl, more preferably -OC(O)C 1-3 alkyl), C 1-8 alkyl (preferably C 1-6 alkyl, more preferably C 1-3 alkyl), or C 1-8 alkoxy (preferably C 1-6 alkoxy, more preferably C 1-3 alkoxy); or R G2a , R G2b together with the attached carbon atom form a 3- to 7-membered saturated or partially unsaturated monocyclic heterocycle or a 3- to 7-membered saturated or partially unsaturated monocyclic ring; wherein the C 1-8 alkyl, C 1-8 alkoxy, 3- to 7-membered saturated or partially unsaturated monocyclic heterocycle and 3- to 7-membered saturated or partially unsaturated monocyclic ring are unsubstituted or substituted by 1, 2 or 3 substituents each independently selected from the group consisting of: deuterium, halogen, cyano, hydroxy, carboxy, C 1-3 alkyl, C 1-3 alkoxy, C 2-4 alkenyl, C 2-4 alkynyl, halo C 1-3 alkyl, halo C 1-3 alkoxy, NR a0 R b0 , -SO 2 C 1-3 alkyl, -S(O)C 1-3 alkyl, -C(O)NR a0 R b0 , -C(O)OC 1-3 alkyl, -OC(O)C 1-3 alkyl, C 3-6 cycloalkyl, C 3-6 cycloalkyloxy, 3- to 6-membered heterocycloalkyl, phenyl, 5- to 6-membered heteroaryl;

[0069] R G2c is hydrogen, C 1-8 alkyl (preferably C 1-6 alkyl, more preferably C 1-3 alkyl), C 3-8 cycloalkyl (preferably C 3-6 cycloalkyl), 3- to 8-membered heterocycloalkyl, -C 1-4 alkyl-3- to 8-membered heterocycloalkyl, -SO 2 C 1-8 alkyl (preferably -SO 2 C 1-6 alkyl, more preferably -SO2 C 1-3 alkyl), -C(O)NR a0 R b0 、-C(O)C 1-8 alkyl (preferably -C(O)C 1-6 alkyl, more preferably -C(O)C 1-3 alkyl), -C(O)OC 1-8 alkyl (preferably -C(O)OC 1-6 alkyl, more preferably -C(O)OC 1-3 alkyl) or C(O)NR a1 R b1 ; wherein said C 1-8 alkyl, C 3-8 cycloalkyl, 3- to 8-membered heterocycloalkyl, -SO 2 C 1-8 alkyl, -C(O)C 1-8 alkyl and -C(O)OC 1-8 alkyl are unsubstituted or substituted with 1, 2 or 3 substituents independently selected from the group consisting of: cyano, hydroxy, carboxy, halogen, acetyl, C 1-3 alkyl, C 1-3 alkoxy, C 2-4 alkenyl, C 2-4 alkynyl, -SO 2 C 1-3 alkyl, -S(O)C 1-3 alkyl, -C(O)NR a0 R b0 、-C(O)OC 1-3 alkyl, -OC(O)C 1-3 alkyl, -C(O)C 1-3 alkyl, -C(O)NR a1 R b1 、NR a1 R b1 、-NR c1 C(O)C 1-3 alkyl, -NR c1 C(O)C 3-6 cycloalkyl, 4- to 6-membered heterocycloalkyl;

[0070] Each R G3a is the same or different, each R G3b is the same or different, and R G3a and R G3b are each independently hydrogen, cyano, hydroxy, carboxy, halogen (preferably fluorine or chlorine), -C(O)OC 1-8 alkyl (preferably -C(O)OC 1-6 alkyl, more preferably -C(O)OC 1-3(alkyl), -OC(O)C 1-8 (alkyl, preferably -OC(O)C 1-6 (alkyl, more preferably -OC(O)C 1-3 (alkyl), C 1-8 (alkyl, preferably C 1-6 (alkyl, more preferably C 1-3 (alkyl) or C 1-8 (alkoxy, preferably C 1-6 (alkoxy, more preferably C 1-3 (alkoxy); or R G3a and R G3b are linked to the same carbon atom, or R G3a and R G3b are linked to different carbon atoms to form a 3- to 7-membered saturated or partially unsaturated monocyclic heterocycle or a 3- to 7-membered saturated or partially unsaturated monocyclic ring; wherein the C 1-8 (alkyl), C 1-8 (alkoxy), 3- to 7-membered saturated or partially unsaturated monocyclic heterocycle, and 3- to 7-membered saturated or partially unsaturated monocyclic ring are unsubstituted or substituted with 1, 2, or 3 substituents each independently selected from the group consisting of deuterium, halogen, cyano, hydroxy, carboxy, C 1-3 (alkyl), C 1-3 (alkoxy), C 2-4 (alkenyl), C 2-4 (alkynyl), halo-C 1-3 (alkyl), halo-C 1-3 (alkoxy), -SO 2 (C 1-3 (alkyl), -S(O)C 1-3 (alkyl), -C(O)NR a0 R b0 (, -C(O)OC 1-3 (alkyl), -OC(O)C 1-3 (alkyl), C 3-6 (cycloalkyl), C 3-6 (cycloalkyloxy), 3- to 6-membered heterocycloalkyl, phenyl, 5- to 6-membered heteroaryl;

[0071] p is 1, 2, or 3;

[0072] R a1 , R b1 are each independently hydrogen, C 1-3 (alkyl), or C 3-6 (cycloalkyl); R c1 is hydrogen or C 1-3 (alkyl);

[0073] R 5a , R a0 , R b0 are as defined in the specification.

[0074] In some embodiments, the structure is the structure represented by formula (d) or formula (e):

[0075]

[0076] wherein R 4 ', R 5 ' are each independently hydrogen, cyano, hydroxy, carboxy, halogen (preferably fluorine or chlorine), -C(O)OC 1-8 alkyl (preferably -C(O)OC 1-6 alkyl, more preferably -C(O)OC 1-3 alkyl), C 1-8 alkyl (preferably C 1-6 alkyl, more preferably C 1-3 alkyl) or C 1-8 alkoxy (preferably C 1-6 alkoxy, more preferably C 1-3 alkoxy); wherein the C 1-8 alkyl, C 1-8 alkoxy is unsubstituted or substituted by 1, 2 or 3 substituents each independently selected from the group consisting of deuterium, halogen, cyano, hydroxy, carboxy, C 1-3 alkyl, C 1-3 alkoxy, C 2-4 alkenyl, C 2-4 alkynyl, halo C 1-3 alkyl, halo C 1-3 alkoxy, NR a0 R b0 2 , -SO C 1-3 alkyl, -S(O)C 1-3 alkyl, -C(O)NR a0 R b0 1-3 , -C(O)OC 1-3 alkyl, -OC(O)C 3-6 alkyl, C 3-6 cycloalkyl, C 1

[0077] X 1 is CR X1a R X1b X1c , NR or O;

[0078] X 2 is CR X2a R X2b ;

[0079] X 3 is a bond or CRX3a R X3b ;

[0080] X 4 is CR X4a R X4b , NR X4c or O;

[0081] wherein R X1a , R X1b , R X2a , R X2b , R X3a , R X3b , R X4a , R X4b are each independently hydrogen, cyano, hydroxy, carboxy, halogen (preferably fluorine or chlorine), -C(O)OC 1-8 alkyl (preferably -C(O)OC 1-6 alkyl, more preferably -C(O)OC 1-3 alkyl), -OC(O)C 1-8 alkyl (preferably -OC(O)C 1-6 alkyl, more preferably -OC(O)C 1-3 alkyl), C 1-8 alkyl (preferably C 1-6 alkyl, more preferably C 1-3 alkyl) or C 1-8 alkoxy (preferably C 1-6 alkoxy, more preferably C 1-3 alkoxy); wherein said C 1-8 alkyl, C 1-8 alkoxy is unsubstituted or substituted by 1, 2 or 3 substituents each independently selected from the group consisting of deuterium, halogen, cyano, hydroxy, carboxy, C 1-3 alkyl, C 1-3 alkoxy, C 2-4 alkenyl, C 2-4 alkynyl, halo C 1-3 alkyl, halo C 1-3 alkoxy, NR a0 R b0 , -SO 2 C 1-3 alkyl, -S(O)C 1-3 alkyl, -C(O)NR a0 R b0 , -C(O)OC 1-3 alkyl, -OC(O)C 1-3 alkyl, C 3-6 cycloalkyl, C 3-6 cycloalkyloxy, 3- to 6-membered heteroalkyl, phenyl, 5- to 6-membered heteroaryl;

[0082] R X1c and R X4c are each independently hydrogen, C 1-8 alkyl (preferably C 1-6 alkyl, more preferably C 1-3 alkyl), C 3-8 cycloalkyl (preferably C 3-6 cycloalkyl), 3- to 8-membered heterocycloalkyl, -C 1-4 alkyl-3- to 8-membered heterocycloalkyl, -SO 2 C 1-8 alkyl (preferably -SO 2 C 1-6 alkyl, more preferably -SO 2 C 1-3 alkyl), -C(O)NR a0 R b0 and -C(O)C 1-8 alkyl (preferably -C(O)C 1-6 alkyl, more preferably -C(O)C 1-3 alkyl), -C(O)OC 1-8 alkyl (preferably -C(O)OC 1-6 alkyl, more preferably -C(O)OC 1-3 alkyl) or C(O)NR a1 R b1 ; where the C 1-8 alkyl, C 3-8 cycloalkyl, 3- to 8-membered heterocycloalkyl, -SO 2 C 1-8 alkyl, -C(O)C 1-8 alkyl and -C(O)OC 1-8 alkyl are unsubstituted or substituted with 1, 2 or 3 substituents independently selected from the group consisting of: cyano, hydroxy, carboxy, halogen, acetyl, C 1-3 alkyl, C 1-3 alkoxy, C 2-4 alkenyl, C 2-4 alkynyl, -SO 2 C 1-3 alkyl, -S(O)C 1-3 alkyl, -C(O)NR a0 R b0 and -C(O)OC 1-3 alkyl, -OC(O)C 1-3 alkyl, -C(O)C 1-3 alkyl, -C(O)NR a1 R b1 and NR a1 R b1 and -NR c1 C(O)C 1-3alkyl, -NR c1 C(O)C 3-6 cycloalkyl, 4- to 6-membered heteroalkyl;

[0083] or R X1a 、R X1b together with the attached carbon atom form a 3- to 7-membered saturated or partially unsaturated monocyclic heterocycle or a 3- to 7-membered saturated or partially unsaturated monocyclic ring; wherein the 3- to 7-membered saturated or partially unsaturated monocyclic heterocycle and the 3- to 7-membered saturated or partially unsaturated monocyclic ring are unsubstituted or substituted with 1, 2, or 3 substituents each independently selected from the group consisting of deuterium, halogen, cyano, hydroxy, carboxy, C 1-3 alkyl, C 1-3 alkoxy, C 2-4 alkenyl, C 2-4 alkynyl, halo C 1-3 alkyl, halo C 1-3 alkoxy, NR a0 R b0 、-SO 2 C 1-3 alkyl, -S(O)C 1-3 alkyl, -C(O)NR a0 R b0 、-C(O)OC 1-3 alkyl, -OC(O)C 1-3 alkyl, C 3-6 cycloalkyl, C 3-6 cycloalkoxy, 3- to 6-membered heteroalkyl, phenyl, 5- to 6-membered heteroaryl;

[0084] or R X2a 、R X2b together with the attached carbon atom form a 3- to 7-membered saturated or partially unsaturated monocyclic heterocycle or a 3- to 7-membered saturated or partially unsaturated monocyclic ring; wherein the 3- to 7-membered saturated or partially unsaturated monocyclic heterocycle and the 3- to 7-membered saturated or partially unsaturated monocyclic ring are unsubstituted or substituted with 1, 2, or 3 substituents each independently selected from the group consisting of deuterium, halogen, cyano, hydroxy, carboxy, C 1-3 alkyl, C 1-3 alkoxy, C 2-4 alkenyl, C 2-4 alkynyl, halo C 1-3 alkyl, halo C 1-3 alkoxy, NR a0 R b0 、-SO 2 C 1-3 alkyl, -S(O)C 1-3 alkyl, -C(O)NR a0 R b0 、-C(O)OC 1-3 alkyl, -OC(O)C1-3 alkyl, C 3-6 cycloalkyl, C 3-6 cycloalkyloxy, 3- to 6-membered heterocycloalkyl, phenyl, 5- to 6-membered heteroaryl;

[0085] or R X3a , R X3b together with the attached carbon atom form a 3- to 7-membered saturated or partially unsaturated monocyclic heterocycle or a 3- to 7-membered saturated or partially unsaturated monocyclic ring; wherein the 3- to 7-membered saturated or partially unsaturated monocyclic heterocycle and the 3- to 7-membered saturated or partially unsaturated monocyclic ring are unsubstituted or substituted with 1, 2, or 3 substituents each independently selected from the group consisting of deuterium, halogen, cyano, hydroxy, carboxy, C 1-3 alkyl, C 1-3 alkoxy, C 2-4 alkenyl, C 2-4 alkynyl, haloC 1-3 alkyl, haloC 1-3 alkoxy, NR a0 R b0 , -SO 2 C 1-3 alkyl, -S(O)C 1-3 alkyl, -C(O)NR a0 R b0 , -C(O)OC 1-3 alkyl, -OC(O)C 1-3 alkyl, C 3-6 cycloalkyl, C 3-6 cycloalkyloxy, 3- to 6-membered heterocycloalkyl, phenyl, 5- to 6-membered heteroaryl;

[0086] or R X4a , R X4b together with the attached carbon atom form a 3- to 7-membered saturated or partially unsaturated monocyclic heterocycle or a 3- to 7-membered saturated or partially unsaturated monocyclic ring; wherein the 3- to 7-membered saturated or partially unsaturated monocyclic heterocycle and the 3- to 7-membered saturated or partially unsaturated monocyclic ring are unsubstituted or substituted with 1, 2, or 3 substituents each independently selected from the group consisting of deuterium, halogen, cyano, hydroxy, carboxy, C 1-3 alkyl, C 1-3 alkoxy, C 2-4 alkenyl, C 2-4 alkynyl, haloC 1-3 alkyl, haloC 1-3 alkoxy, NR a0 R b0 , -SO 2 C 1-3 alkyl, -S(O)C 1-3 alkyl, -C(O)NR a0 R b0 , -C(O)OC1-3 alkyl, -OC(O)C 1-3 alkyl, C 3-6 cycloalkyl, C 3-6 cycloalkyloxy, 3- to 6-membered heterocycloalkyl, phenyl, 5- to 6-membered heteroaryl;

[0087] or R X4c joins with R X3a to form a fused 4- to 7-membered saturated or partially unsaturated monocyclic heterocycle; wherein the 4- to 7-membered saturated or partially unsaturated monocyclic heterocycle is unsubstituted or substituted with 1, 2, or 3 substituents each independently selected from the group consisting of deuterium, halogen, cyano, hydroxy, carboxy, C 1-3 alkyl, C 1-3 alkoxy, C 2-4 alkenyl, C 2-4 alkynyl, haloC 1-3 alkyl, haloC 1-3 alkoxy, NR a0 R b0 、-SO 2 C 1-3 alkyl, -S(O)C 1-3 alkyl, -C(O)NR a0 R b0 、-C(O)OC 1-3 alkyl, -OC(O)C 1-3 alkyl, C 3-6 cycloalkyl, C 3-6 cycloalkyloxy, 3- to 6-membered heterocycloalkyl, phenyl, 5- to 6-membered heteroaryl;

[0088] or R X1c joins with R X2a to form a fused 4- to 7-membered saturated or partially unsaturated monocyclic heterocycle; wherein the 4- to 7-membered saturated or partially unsaturated monocyclic heterocycle is unsubstituted or substituted with 1, 2, or 3 substituents each independently selected from the group consisting of deuterium, halogen, cyano, hydroxy, carboxy, C 1-3 alkyl, C 1-3 alkoxy, C 2-4 alkenyl, C 2-4 alkynyl, haloC 1-3 alkyl, haloC 1-3 alkoxy, NR a0 R b0 、-SO 2 C 1-3 alkyl, -S(O)C 1-3 alkyl, -C(O)NR a0 R b0 、-C(O)OC 1-3 alkyl, -OC(O)C 1-3 alkyl, C 3-6Cycloalkyl, C 3-6 Cycloalkyloxy, 3- to 6-membered heterocycloalkyl, phenyl, 5- to 6-membered heteroaryl;

[0089] R a1 、R b1 are each independently hydrogen, C 1-3 alkyl or C 3-6 cycloalkyl; R c1 is hydrogen or C 1-3 alkyl;

[0090] R 1 、R 2 、R 3 、R a0 、R b0 are as defined in the above specification.

[0091] In some embodiments, X 1 is NR X1c or O; X 2 is CR X2a R X2b ; X 3 is a bond or CR X3a R X3b ; X 4 is NR X4c or O.

[0092] In some embodiments, X 1 is NR X1c ; X 2 is CR X2a R X2b ; X 3 is a bond or CR X3a R X3b ; X 4 is O.

[0093] In some embodiments, X 1 is O; X 2 is CR X2a R X2b ; X 3 is a bond or CR X3a R X3b ; X 4 is NR X4c or O.

[0094] In some embodiments, in the above structural formulas, the 3- to 7-membered saturated or partially unsaturated monocyclic heterocycles in each group are 4- to 6-membered saturated or partially unsaturated monocyclic heterocycles, each independently selected from: azetidine, oxetane, tetrahydrofuran ring, tetrahydrothiophene ring, pyrrolidine ring, piperidine ring, piperazine ring, morpholine ring, thiomorpholine ring, thiomorpholine-1,1-dioxide, tetrahydropyran ring, 1,2-dihydroazetadiene ring, 1,2-dihydrooxetadiene ring, 2,5-dihydro-1H-pyrrole ring, 2,5-dihydrofuran ring, 2,3-dihydrofuran ring, 2,3-dihydro-1H-pyrrole ring, 3,4-dihydro-2H-pyran ring, 1,2,3,4-tetrahydropyridine ring, 3,6-dihydro-2H-pyran ring, and 1,2,3,6-tetrahydropyridine ring; more preferably 4- to 6-membered saturated monocyclic heterocycles.

[0095] In some embodiments, in the above structural formulas, the 3- to 7-membered saturated or partially unsaturated monocyclic rings in each group are 3- to 6-membered saturated or partially unsaturated monocyclic rings, each independently selected from: cyclopropyl ring, cyclobutyl ring, cyclopentyl ring, cyclopentenyl ring, cyclohexyl ring, cyclohexenyl ring, and cyclohexadienyl ring; more preferably 3- to 6-membered saturated monocyclic rings.

[0096] In some embodiments, in the above structural formulas, the 5- to 6-membered heteroaryl groups in each group are each independently selected from: thienyl, furyl, thiazolyl, imidazolyl, oxazolyl, pyrrolyl, pyrazolyl, triazolyl, 1,2,3-triazolyl, 1,2,4-triazolyl, 1,2,5-triazolyl, 1,3,4-triazolyl, tetrazolyl, isoxazolyl, oxadiazolyl, 1,2,3-oxadiazolyl, 1,2,4-oxadiazolyl, 1,2,5-oxadiazolyl, 1,3,4-oxadiazolyl, thiadiazolyl, pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl.

[0097] In some embodiments, in the above structural formulas, the 3- to 6-membered heterocyclic alkyl groups in each group are 4- to 6-membered heterocyclic alkyl groups, each independently selected from: azetidinyl, oxetanyl, tetrahydrofuryl, tetrahydrothiophenyl, pyrrolidinyl, oxazolidinyl, dioxolanyl, piperidinyl, piperazinyl, morpholinyl, dioxanyl, thiomorpholinyl, thiomorpholine-1,1-dioxide, tetrahydropyranyl, pyrrolidin-2-one group, dihydrofuran-2(3H)-one group, morpholin-3-one group, piperazine-2-one group, piperidin-2-one group.

[0098] In some embodiments, in the above structural formulas, the 5- to 8-membered saturated or partially unsaturated monocyclic heterocycles in each group are each independently selected from: imidazolidine, hexahydropyrimidine ring, 1,3-diazepine ring, 2,3-dihydro-1H-imidazole ring, 1,2,3,4-tetrahydropyrimidine ring, 2,3,4,5-tetrahydro-1H-1,3-diazepine ring, 2,3,4,7-tetrahydro-1H-1,3-diazepine ring.

[0099] In some embodiments, in the above structural formulas, the 5- to 8-membered heteroalkenyl rings in each group are selected from: 4,5-dihydro-1H-imidazole ring, 1,4,5,6-tetrahydropyrimidine ring, 3,4,7,8-tetrahydro-2H-1,4,6-oxadiazine ring, 1,6-dihydropyrimidine ring, 4,5,6,7-tetrahydro-1H-1,3-diazepine ring, 2,5,6,7-tetrahydro-1,3,5-oxadiazepine ring, 4,5-dihydrooxazole ring, 5,6-dihydro-4H-1,3-oxazine ring, 6H-1,3-oxazine ring, 4,5,6,7-tetrahydro-1,3-oxazepane, 6,7-dihydro-2H-1,5,3-dioxazepane.

[0100] In some embodiments, R 4 and R 5 are connected to form a fused 3- to 7-membered saturated or partially unsaturated monocyclic heterocycle which is a fused 4- to 6-membered saturated monocyclic heterocycle, selected from: azetidine, oxetane, tetrahydrofuran ring, tetrahydrothiophene ring, tetrahydropyrrole ring, piperidine ring, oxazolidine, piperazine ring, dioxolane, 1,4-dioxane, morpholine ring, thiomorpholine ring, tetrahydropyran ring, morpholin-3-one ring, piperazin-2-one ring, piperidin-2-one ring.

[0101] In some embodiments, R 4 and R 5 are connected to form a fused 3- to 7-membered saturated or partially unsaturated monocyclic ring which is a fused 3- to 6-membered saturated monocyclic ring, selected from: cyclopropyl ring, cyclobutyl ring, cyclopentyl ring, cyclohexyl ring.

[0102] In some embodiments, any two R s4 connected on the same ring atom or different ring atoms to form a 3- to 6-membered saturated or partially unsaturated monocyclic heterocycle which is a 4- to 6-membered saturated monocyclic heterocycle, selected from: azetidine, oxetane, tetrahydrofuran ring, tetrahydrothiophene ring, tetrahydropyrrole ring, piperidine ring, piperazine ring, morpholine ring, thiomorpholine ring, thiomorpholine-1,1-dioxide, tetrahydropyran ring.

[0103] In some embodiments, any two R s4The 3- to 6-membered saturated or partially unsaturated monocyclic ring formed by connection is a 3- to 6-membered saturated monocyclic ring, selected from: cyclopropyl ring, cyclobutyl ring, cyclopentyl ring, cyclohexyl ring.

[0104] In some embodiments, R a0 , R b0 The 4- to 6-membered saturated monocyclic heterocycle formed together with the connected nitrogen atom is selected from: azetidine, oxetane, tetrahydrofuran ring, tetrahydrothiophene ring, pyrrolidine ring, piperidine ring, piperazine ring, morpholine ring, thiomorpholine ring, thiomorpholine-1,1-dioxide, tetrahydropyran ring.

[0105] In some embodiments, formula (a) is selected from the following structures:

[0106]

[0107] In some embodiments, formula (a) is selected from the following structures:

[0108]

[0109] In some embodiments, formula (b1) is selected from the following structures:

[0110]

[0111] In some embodiments, formula (b2) is selected from the following structures:

[0112]

[0113] In some embodiments, formula (c1) is selected from the following structures:

[0114]

[0115] In some embodiments, when Q is a 5- to 10-membered heteroaryl group, the 5- to 10-membered heteroaryl group is a 5- to 6-membered heteroaryl group or an 8- to 10-membered heteroaryl group.

[0116] In some embodiments, the 5- to 6-membered heteroaryl group is selected from: thienyl, furyl, thiazolyl, isothiazolyl, imidazolyl, oxazolyl, pyrrolyl, pyrazolyl, triazolyl, 1,2,3-triazolyl, 1,2,4-triazolyl, 1,2,5-triazolyl, 1,3,4-triazolyl, tetrazolyl, isoxazolyl, oxadiazolyl, 1,2,3-oxadiazolyl, 1,2,4-oxadiazolyl, 1,2,5-oxadiazolyl, 1,3,4-oxadiazolyl, thiadiazolyl, pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, triazinyl and tetrazinyl.

[0117] In some embodiments, the 8- to 10-membered heteroaryl is selected from: indolyl, isoindolyl, indazolyl, benzotriazolyl, benzothienyl, isobenzothienyl, benzofuranyl, isobenzofuranyl, benzimidazolyl, benzoxazolyl, benzisoxazolyl, benzoxadiazolyl, benzothiazolyl, benzisothiazolyl, benzothiadiazolyl, indolizinyl, purinyl, pyrido[3,2-d]pyrimidinyl, pyrido[2,3-d]pyrimidinyl, pyrido[3,4-d]pyrimidinyl, pyrido[4,3-d]pyrimidinyl, 1,8-naphthyridinyl, 1,7-naphthyridinyl, 1,6-naphthyridinyl, 1,5-naphthyridinyl, pteridinyl, quinolinyl, isoquinolinyl, cinnolinyl, quinoxalinyl, phthalazinyl, and quinazolinyl.

[0118] In some embodiments, the 5- to 6-membered heteroaryl is selected from:

[0119]

[0120] The above 5- to 6-membered heteroaryl is unsubstituted or substituted with 1, 2, or 3 substituents each independently selected from the group consisting of: deuterium, halogen, cyano, hydroxy, carboxy, C 1-3 alkyl, C 1-3 alkoxy, C 2-4 alkenyl, C 2-4 alkynyl, haloC 1-3 alkyl, haloC 1-3 alkoxy, NR a0 R b0 、-SO 2 C 1-3 alkyl, -S(O)C 1-3 alkyl, -C(O)NR a0 R b0 、-C(O)OC 1-3 alkyl, -OC(O)C 1-3 alkyl, C 3-6 cycloalkyl, C 3-6 cycloalkyloxy, 3- to 6-membered heterocycloalkyl, phenyl, 5- to 6-membered heteroaryl; R a0 、R b0 as defined in the above specification.

[0121] In some embodiments, the 5- to 6-membered heteroaryl is selected from:

[0122]

[0123] In some embodiments, the 8- to 10-membered heteroaryl is selected from:

[0124] In some embodiments, Q is selected from the following group of structures:

[0125]

[0126]

[0127] In some embodiments, Q is selected from the following group of structures:

[0128]

[0129] In some embodiments, formula (e) is selected from the following structures:

[0130]

[0131] wherein R 1 , R 2 , R 3 are as defined in the above specification. In some embodiments, R 1 is hydrogen or methoxy.

[0132] In some embodiments, R 2 is hydrogen.

[0133] In some embodiments, R 3 is fluorine.

[0134] In some embodiments, R 4 , R 5 are each independently hydrogen.

[0135] In some embodiments, the structure is selected from the following structures:

[0136]

[0137] In some embodiments, the compound of formula (I) is any one of the following compounds:

[0138]

[0139] In some embodiments, the compound of formula (I) is any one of the following compounds:

[0140]

[0141]

[0142]

[0143] In some embodiments, the compound of formula (I) is any one of the following compounds:

[0144]

[0145]

[0146] In some embodiments, the compound of formula (I) is any one of the following compounds:

[0147]

[0148] The second aspect of the present invention provides a pharmaceutical composition, which comprises the compound described in the first aspect of the present invention, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, or a solvate thereof, or a prodrug thereof, and a pharmaceutically acceptable carrier.

[0149] The third aspect of the present invention provides the use of the compound described in the first aspect of the present invention, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, or a solvate thereof, or a prodrug thereof, and the pharmaceutical composition described in the second aspect of the present invention in the preparation of a medicament for treating a disease associated with or mediated by CDK9 activity. The disease is preferably a hyperproliferative disease, a virus-induced infectious disease and / or a cardiovascular disease, more preferably a hyperproliferative disease.

[0150] The fourth aspect of the present invention provides a method for treating a disease associated with or mediated by CDK9 activity, the method comprising administering to a patient an effective amount of the compound described in the first aspect of the present invention, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, or a solvate thereof, or a prodrug thereof, or the pharmaceutical composition as described in the second aspect of the present invention. In certain embodiments, the disease associated with CDK9 activity is a hyperproliferative disease, especially cancer, such as solid tumors and hematological tumors.

[0151] In the present invention, diseases associated with or mediated by CDK9 activity include diseases associated with or involving CDK9 activity (such as overactivity of CDK9), and conditions associated with these diseases. The overactivity of CDK9 refers to an increased CDK9 enzyme activity compared to normal non-diseased cells, or it refers to an increased CDK9 activity that leads to unwanted cell proliferation, or reduced or insufficient programmed cell death (apoptosis), or it refers to a mutation that results in constitutive activation of CDK9. Hyperproliferative diseases include diseases involving unwanted or uncontrolled proliferation of cells, and include diseases involving reduced or insufficient programmed cell death (apoptosis). The compounds of the present invention can be used for preventing, inhibiting, blocking, reducing, lowering, controlling, etc. cell proliferation and / or cell division, and / or inducing apoptosis. The method includes administering to a subject in need thereof (including mammals, including humans) an amount of the compound of the present invention or a pharmaceutically acceptable salt, hydrate or solvate thereof effective for treating or preventing the disease.

[0152] Hyperproliferative diseases in the context of the present invention include, but are not limited to, for example, angiogenic or vasculoproliferative disorders, mesangial cell proliferative diseases, and solid tumors such as cancers of the breast, respiratory tract, brain, reproductive organs, digestive tract, urinary tract, eye, liver, skin, head and neck, thyroid, parathyroid, and their distant metastases. Those diseases also include lymphomas, sarcomas, and leukemias. In some embodiments, the cancers are selected from pancreatic cancer, breast cancer, ovarian cancer, cervical cancer, leukemia.

[0153] It should be understood that within the scope of the present invention, each of the above technical features of the present invention and the technical features specifically described below (such as in the examples) can be combined with each other to form new or preferred technical solutions. Due to space limitations, they will not be elaborated one by one here. BRIEF DESCRIPTION OF THE DRAWINGS

[0154] Figure 1 Graph showing the effect of compound Z-1 on the tumor volume of animals (the abscissa is the number of days (days), and the ordinate is the tumor volume TV (mm 3 )

[0155] Figure 2 Relative tumor growth rate of compound Z-1 (the abscissa is the number of days (days), and the ordinate is the relative tumor growth rate T / C (%));

[0156] Figure 3 Graph showing the effect of compound Z-1 on the body weight change of tumor-bearing animals (the abscissa is the number of days (days), and the ordinate is the body weight change rate BW (%) of the animals). DETAILED DESCRIPTION OF THE INVENTION

[0157] Through extensive and in-depth research, the present inventors unexpectedly discovered such substituted bis(pyridin-2-yl)amine derivatives, which have significant CDK9 selective inhibitory activity and excellent in vivo pharmacokinetic activity. Therefore, this series of compounds is expected to be developed into drugs for treating and / or preventing diseases related to or mediated by CDK9 activity. On this basis, the inventors completed the present invention.

[0158] TERM DEFINITION

[0159] To be able to more clearly understand the technical content of the present invention, the terms of the present invention are further described below.

[0160] "Alkyl" refers to straight-chain and branched-chain saturated aliphatic hydrocarbon groups. "C 1-8 alkyl" refers to an alkyl group having 1 to 8 carbon atoms, preferably C 1-6 alkyl, more preferably C 1-3Alkyl; Non-limiting examples of alkyl include: methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, 1,1-dimethylpropyl, 1,2-dimethylpropyl, 2,2-dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, 1-ethyl-2-methylpropyl, 1,1,2-trimethylpropyl, 1,1-dimethylbutyl, 1,2-dimethylbutyl, 2,2-dimethylbutyl, 1,3-dimethylbutyl, 2-ethylbutyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 2,3-dimethylbutyl, n-heptyl, 2-methylhexyl, 3-methylhexyl, 4-methylhexyl, 5-methylhexyl, 2,3-dimethylpentyl, 2,4-dimethylpentyl, 2,2-dimethylpentyl, 3,3-dimethylpentyl, 2-ethylpentyl, 3-ethylpentyl, n-octyl, 2,3-dimethylhexyl, 2,4-dimethylhexyl, 2,5-dimethylhexyl, 2,2-dimethylhexyl, 3,3-dimethylhexyl, 4,4-dimethylhexyl, 2-ethylhexyl, 3-ethylhexyl, 4-ethylhexyl, 2-methyl-2-ethylpentyl, 2-methyl-3-ethylpentyl, n-nonyl, 2-methyl-2-ethylhexyl, 2-methyl-3-ethylhexyl, 2,2-diethylpentyl, n-decyl, 3,3-diethylhexyl, 2,2-diethylhexyl, and various branched isomers thereof are more preferred.

[0161] "Alkenyl" refers to a straight-chain or branched-chain unsaturated aliphatic hydrocarbon group having one or more carbon-carbon double bonds (C═C), "C" 2-8 Alkenyl" refers to an alkenyl having 2 to 8 carbon atoms, preferably C 2-6 Alkenyl, more preferably C 2-4 Alkenyl, with a similar definition; non-limiting examples include vinyl, propenyl, isopropenyl, n-butenyl, isobutenyl, pentenyl, hexenyl, etc.

[0162] "Alkynyl" refers to a straight-chain and branched-chain unsaturated aliphatic hydrocarbon group having one or more carbon-carbon triple bonds, "C" 2-8 Alkynyl" refers to an alkynyl having 2 to 8 carbon atoms, preferably C 2-6 Alkynyl, more preferably C 2-4 Alkynyl, with a similar definition; non-limiting examples include ethynyl, propynyl, n-butynyl, isobutynyl, pentynyl, hexynyl, etc.

[0163] "Cycloalkyl" and "cycloalkyl ring" are used interchangeably and both refer to a saturated monocyclic, bicyclic or polycyclic cyclic hydrocarbon group, which group may be fused to an aryl or heteroaryl group. The cycloalkyl ring may be optionally substituted. In certain embodiments, the cycloalkyl ring contains one or more carbonyl groups, such as an oxo group. "C" 3-8"Cycloalkyl" refers to a monocyclic cycloalkyl group having 3 to 8 carbon atoms. Non-limiting examples of cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclobutanone, cyclopentanone, cyclopentane-1,3-dione, etc. Preferably C 3-6 cycloalkyl, including cyclopropyl, cyclobutyl, cyclopentyl and cyclohexyl. "C 8-10 "Cycloalkyl" refers to a fused bicyclic cycloalkyl group having 8 to 10 ring atoms, C 8-10 Non-limiting examples of cycloalkyl groups include

[0164] "Cycloalkenyl" and "cycloalkenyl ring" are used interchangeably and both refer to a monocyclic, bicyclic or polycyclic cycloalkyl group containing one or more carbon-carbon double bonds in the ring, which group may be fused to an aryl or heteroaryl group. The cycloalkenyl ring may be optionally substituted. In certain embodiments, the cycloalkenyl ring contains one or more carbonyl groups, such as oxo groups. "C 3-8 "Cycloalkenyl" refers to a monocyclic cycloalkenyl group having 3 to 8 carbon atoms. Preferably C 3-6 cycloalkenyl. Non-limiting examples of cycloalkenyl groups include cyclobutenyl, cyclopentenyl, cyclohexenyl, cyclohexadienyl, cycloheptatrienyl, cyclopentyl-2-en-1-one, cyclohexyl-2,5-dien-1-one, cyclohexyl-2-en-1-one, cyclohex-2-ene-1,4-dione, etc.

[0165] "Heterocycloalkyl" and "heterocycloalkyl ring" are used interchangeably and each refers to a cycloalkyl group containing at least one heteroatom selected from nitrogen, oxygen and sulfur, and the group may be fused to an aryl or heteroaryl group. The heterocycloalkyl ring may be optionally substituted. In certain embodiments, the heterocycloalkyl ring contains one or more carbonyl or thiocarbonyl groups, such as groups containing oxo and thio. "3- to 8-membered heterocycloalkyl" refers to a monocyclic cyclic hydrocarbon group having 3 to 8 ring atoms, wherein 1, 2 or 3 ring atoms are heteroatoms selected from nitrogen, oxygen and sulfur, preferably 4- to 8-membered heterocycloalkyl. More preferably, it is a 3- to 6-membered heterocycloalkyl having 3 to 6 ring atoms, wherein 1 or 2 ring atoms are heteroatoms selected from nitrogen, oxygen and sulfur. More preferably, it is a 4- to 6-membered heterocycloalkyl having 4 to 6 ring atoms, wherein 1 or 2 ring atoms are heteroatoms selected from nitrogen, oxygen and sulfur. Non-limiting examples of monocyclic heterocycloalkyl include aziridinyl, oxiranyl, azetidinyl, oxetanyl, tetrahydrofuranyl, tetrahydrothienyl, pyrrolidinyl, oxazolidinyl, dioxolanyl, piperidinyl, piperazinyl, morpholinyl, dioxanyl, thiomorpholinyl, thiomorpholine-1,1-dioxide, tetrahydropyranyl, azetidin-2-one, oxetane-2-one, dihydrofuran-2(3H)-one, pyrrolidin-2-one, pyrrolidine-2,5-dione, dihydrofuran-2,5-dione, piperidin-2-one, tetrahydro-2H-pyran-2-one, piperazine-2-one, morpholin-3-one, etc. "6- to 12-membered heterocycloalkyl" and "6- to 12-membered fused heterocycloalkyl" are used interchangeably and refer to a fused bicyclic cyclic hydrocarbon group having 6 to 12 ring atoms, wherein 1, 2 or 3 ring atoms are heteroatoms selected from nitrogen, oxygen and sulfur. "8- to 10-membered heterocycloalkyl" and "8- to 10-membered fused heterocycloalkyl" are used interchangeably and refer to a fused bicyclic cyclic hydrocarbon group having 8 to 10 ring atoms, wherein 1, 2 or 3 ring atoms are heteroatoms selected from nitrogen, oxygen and sulfur. Non-limiting examples of bicyclic heterocycloalkyl include hexahydro-1H-furo[3,4-c]pyrrole, octahydro-1H-cyclopenta[c]pyridine, hexahydro-1H-pyrrolo[2,1-c][1,4]oxazine, octahydropyrrolo[1,2-a]pyrazine, hexahydropyrrolo[1,2-a]pyrazin-4(1H)-one, octahydrocyclopenta[c]pyrrole, etc. In a fused bicyclic heterocycloalkyl containing one or more nitrogen atoms, the point of attachment may be a carbon or nitrogen atom as long as the valence allows. The bicyclic heterocycloalkyl system may include one or more heteroatoms in one or both rings.

[0166] "Heterocyclenyl" and "heterocyclenyl ring" are used interchangeably and refer to a heterocycloalkyl group containing one or more carbon-carbon double bonds or carbon-nitrogen double bonds within the ring, but are not intended to include a heteroaryl moiety as defined herein. The group may be fused to an aryl or heteroaryl group. The heterocyclenyl ring may be optionally substituted. In certain embodiments, the heterocyclenyl ring contains one or more carbonyl or thiocarbonyl groups, such as groups containing oxo and thio. "5- to 8-membered heterocyclenyl ring" refers to a heterocyclenyl ring having 5 to 8 ring atoms, wherein 1, 2, or 3 ring atoms are heteroatoms selected from nitrogen, oxygen, and sulfur. Preferably, it is a 5- to 6-membered heterocyclenyl ring. Non-limiting examples of the heterocyclenyl ring include 4,5-dihydro-1H-imidazole ring, 1,4,5,6-tetrahydropyrimidine ring, 3,4,7,8-tetrahydro-2H-1,4,6-oxadiazine ring, 1,6-dihydropyrimidine ring, 4,5,6,7-tetrahydro-1H-1,3-diazepine ring, 2,5,6,7-tetrahydro-1,3,5-oxadiazepine ring.

[0167] "Aryl" and "aryl ring" are used interchangeably and both refer to a fully carbon monocyclic or fused polycyclic (i.e., rings sharing adjacent carbon atom pairs) group having a conjugated π-electron system, and the group may be fused to a cycloalkyl ring, heterocycloalkyl ring, cycloalkenyl ring, heterocyclenyl ring, or heteroaryl group. "C 6-10 aryl" refers to a monocyclic or bicyclic aryl group having 6 to 10 carbon atoms, and non-limiting examples of the aryl group include phenyl, naphthyl, etc.

[0168] "Heteroaryl" and "heteroaryl ring" are used interchangeably and each refers to a group of a monocyclic, bicyclic or polycyclic 4n+2 aromatic ring system having ring carbon atoms and ring heteroatoms (e.g., having 6 or 10 π electrons shared in a cyclic arrangement), wherein each heteroatom is independently selected from nitrogen, oxygen and sulfur. In the present invention, heteroaryl also includes a ring system in which the above heteroaryl ring is fused with one or more cycloalkyl rings, heterocycloalkyl rings, cycloalkenyl rings, heterocycloalkenyl rings or aromatic rings. The heteroaryl ring may be optionally substituted. "5- to 10-membered heteroaryl" refers to a monocyclic or bicyclic heteroaryl having 5 to 10 ring atoms, wherein 1, 2, 3 or 4 ring atoms are heteroatoms. "5- to 6-membered heteroaryl" refers to a monocyclic heteroaryl having 5 to 6 ring atoms, wherein 1, 2, 3 or 4 ring atoms are heteroatoms, and non-limiting examples include thienyl, furyl, thiazolyl, isothiazolyl, imidazolyl, oxazolyl, pyrrolyl, pyrazolyl, triazolyl, 1,2,3-triazolyl, 1,2,4-triazolyl, 1,2,5-triazolyl, 1,3,4-triazolyl, tetrazolyl, isoxazolyl, oxadiazolyl, 1,2,3-oxadiazolyl, 1,2,4-oxadiazolyl, 1,2,5-oxadiazolyl, 1,3,4-oxadiazolyl, thiadiazolyl, pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, triazinyl, tetrazinyl. "8- to 10-membered heteroaryl" refers to a bicyclic heteroaryl having 8 to 10 ring atoms, wherein 1, 2, 3 or 4 ring atoms are heteroatoms, and non-limiting examples include indolyl, isoindolyl, indazolyl, benzotriazolyl, benzothienyl, isobenzothienyl, benzofuryl, isobenzofuryl, benzimidazolyl, benzoxazolyl, benzisoxazolyl, benzoxadiazolyl, benzothiazolyl, benzisothiazolyl, benzothiadiazolyl, indolizinyl, purinyl, pyrido[3,2-d]pyrimidinyl, pyrido[2,3-d]pyrimidinyl, pyrido[3,4-d]pyrimidinyl, pyrido[4,3-d]pyrimidinyl, 1,8-naphthyridinyl, 1,7-naphthyridinyl, 1,6-naphthyridinyl, 1,5-naphthyridinyl, pteridinyl, quinolinyl, isoquinolinyl, cinnolinyl, quinoxalinyl, phthalazinyl and quinazolinyl. "Heteroatom" refers to nitrogen, oxygen or sulfur. In a heteroaryl containing one or more nitrogen atoms, the point of attachment may be a carbon or nitrogen atom as long as the valence allows. The bicyclic heteroaryl system may include one or more heteroatoms in one or two rings.

[0169] "Fused" refers to a structure in which two or more rings share one or more bonds.

[0170] "Alkoxy" means -O-alkyl, wherein alkyl is defined as above. Preferably C 1-8 alkoxy, more preferably C 1-6 alkoxy, most preferably C 1-3 alkoxy. Non-limiting examples include methoxy, ethoxy, n-propoxy, isopropoxy, butoxy, tert-butoxy, isobutoxy, pentyloxy, etc.

[0171] "Cycloalkyloxy" means -O-cycloalkyl, where cycloalkyl is defined as above. Preferably C 3-8 cycloalkyloxy, more preferably C 3-6 cycloalkyloxy. Non-limiting examples include cyclopropyloxy, cyclobutyloxy, cyclopentyloxy, cyclohexyloxy, etc.

[0172] "A bond" means that the two groups connected by it are connected by a covalent bond.

[0173] "Halogen" means fluorine, chlorine, bromine or iodine.

[0174] "Halogenated" means that one or more (such as 1, 2, 3, 4 or 5) hydrogens in the group are replaced by halogen.

[0175] "Amino" means NH 2 ,"Cyano" means CN, "Nitro" means NO 2 ,"Benzyl" means -CH 2 -phenyl, "Oxo group" means =O, "Carboxyl group" means -C(O)OH, "Acetyl group" means -C(O)CH 3 ,"Hydroxymethyl" means -CH 2 OH, "Hydroxyethyl" means -CH 2 CH 2 OH or -CHOHCH 3 ,"Hydroxyl" means -OH, "Thiol" means SH, "Sub-cyclopropyl" structure is:

[0176] "Saturated or partially unsaturated monocyclic" refers to a saturated or partially unsaturated all-carbon monocyclic system, where "partially unsaturated" means a ring portion including at least one double bond or triple bond, and "partially unsaturated" is intended to cover rings with multiple unsaturated sites, but is not intended to include aryl or heteroaryl moieties as defined herein. In certain embodiments, the saturated or partially unsaturated monocyclic contains one or more carbonyl groups, such as an oxo group. "3- to 7-membered saturated or partially unsaturated monocyclic" has 3 to 7 ring carbon atoms, preferably a saturated or partially unsaturated monocyclic having 3 to 6 ring carbon atoms, more preferably a saturated monocyclic having 3 to 6 ring carbon atoms. Non-limiting examples of saturated or partially unsaturated monocyclics include cyclopropyl ring, cyclobutyl ring, cyclopentyl ring, cyclopentenyl ring, cyclohexyl ring, cyclohexenyl ring, cyclohexadienyl ring, cycloheptyl ring, cycloheptatrienyl ring, cyclopentanone ring, cyclopentane-1,3-dione ring, etc.

[0177] "Saturated or partially unsaturated monocyclic heterocycle" means that 1, 2 or 3 ring carbon atoms in a saturated or partially unsaturated monocyclic are selected from nitrogen, oxygen or S(O) t(wherein t is an integer from 0 to 2) and the remaining ring atoms are carbon, provided that the ring moiety is not -O-O-, -O-S- or -S-S-. "3- to 7-membered saturated or partially unsaturated monocyclic heterocycle" has 3 to 7 ring atoms, wherein 1, 2 or 3 ring atoms are the above-mentioned heteroatoms. Preferably, it is a 3- to 6-membered saturated or partially unsaturated monocyclic heterocycle having 3 to 6 ring atoms, wherein 1 or 2 ring atoms are the above-mentioned heteroatoms, more preferably a 5- to 6-membered saturated or partially unsaturated monocyclic heterocycle having 5 to 6 ring atoms, wherein 1 or 2 ring atoms are the above-mentioned heteroatoms, and most preferably a 5- or 6-membered saturated monocyclic heterocycle. Non-limiting examples of saturated monocyclic heterocycles include an epoxypropane ring, an azetidine ring, an oxetane ring, a tetrahydrofuran ring, a tetrahydrothiophene ring, a pyrrolidine ring, a piperidine ring, a pyrroline ring, an oxazolidine ring, a piperazine ring, a dioxolane ring, a dioxane ring, a morpholine ring, a thiomorpholine ring, thiomorpholine-1,1-dioxide, a tetrahydropyran ring, an azetidin-2-one ring, an oxetane-2-one ring, a pyrrolidin-2-one ring, a pyrrolidine-2,5-dione ring, a piperidin-2-one ring, a dihydrofuran-2(3H)-one ring, a dihydrofuran-2,5-dione ring, a tetrahydro-2H-pyran-2-one ring, a piperazine-2-one ring, a morpholin-3-one ring. Non-limiting examples of partially unsaturated monocyclic heterocycles include 1,2-dihydroazete ring, 1,2-dihydrooxete ring, 2,5-dihydro-1H-pyrrole ring, 2,5-dihydrofuran ring, 2,3-dihydrofuran ring, 2,3-dihydro-1H-pyrrole ring, 3,4-dihydro-2H-pyran ring, 1,2,3,4-tetrahydropyridine ring, 3,6-dihydro-2H-pyran ring, 1,2,3,6-tetrahydropyridine ring, 4,5-dihydro-1H-imidazole ring, 1,4,5,6-tetrahydropyrimidine ring, 3,4,7,8-tetrahydro-2H-1,4,6-oxadiazine ring, 1,6-dihydropyrimidine ring, 4,5,6,7-tetrahydro-1H-1,3-diazepine ring, 2,5,6,7-tetrahydro-1,3,5-oxadiazepine ring, etc.

[0178] "Substituted" means that one or more hydrogen atoms in the group, preferably 1 to 5 hydrogen atoms, are independently replaced by the corresponding number of substituents, more preferably 1 to 3 hydrogen atoms are independently replaced by the corresponding number of substituents. It goes without saying that the substituents are only in their possible chemical positions, and those skilled in the art can determine (by experiment or theory) what is possible or impossible to substitute without undue effort. For example, an amino or hydroxyl group having a free hydrogen may be unstable when combined with a carbon atom having an unsaturated (such as olefinic) bond.

[0179] Unless otherwise defined, the "substituents independently selected from... respectively" as used in the present invention means that when more than one hydrogen on the group is replaced by substituents, the types of the substituents may be the same or different, and the substituents selected are of independent types respectively.

[0180] Unless otherwise defined, the phrase "… the same or different and each independently is …" as used in the present invention means that when there is more than one identical substituent group in the general formula, the groups may be the same or different and are each independent species. For example, when L is (CR e R f ) s , when s is 2, i.e., L is (CR e R f )-(CR e R f ), the two R e or R f may be the same or different and are each independent species.

[0181] Unless otherwise defined, any group herein may be substituted or unsubstituted. When the above groups are substituted, the substituents are preferably 1 to 5 of the following groups, independently selected from cyano, halogen (preferably fluorine or chlorine), C 1-8 alkyl (preferably C 1-6 alkyl, more preferably C 1-3 alkyl), C 1-8 alkoxy (preferably C 1-6 alkoxy, more preferably C 1-3 alkoxy), halo C 1-8 alkyl (preferably halo C 1-6 alkyl, more preferably halo C 1-3 alkyl), C 3-8 cycloalkyl (preferably C 3-6 cycloalkyl), halo C 1-8 alkoxy (preferably halo C 1-6 alkoxy, more preferably halo C 1-3 alkoxy), C 1-8 alkyl-substituted amino, halo C 1-8 alkyl-substituted amino, acetyl, hydroxy, hydroxymethyl, hydroxyethyl, carboxyl, nitro, C 6-10 aryl (preferably phenyl), C 3-8 cycloalkyloxy (preferably C 3-6 cycloalkyloxy), C 2-8 alkenyl (preferably C 2-6 alkenyl, more preferably C 2-4 alkenyl), C 2-8 alkynyl (preferably C 2-6 alkynyl, more preferably C 2-4 alkynyl), -CONR a0 R b0 , -C(O)OC 1-10 alkyl (preferably -C(O)OC 1-6 alkyl, more preferably -C(O)OC 1-3(alkyl), -CHO, -OC(O)C 1-10 (preferably -OC(O)C 1-6 (alkyl, more preferably -OC(O)C 1-3 (alkyl), -SO 2 C 1-10 (preferably -SO 2 C 1-6 (alkyl, more preferably -SO 2 C 1-3 (alkyl), -SO 2 C 6-10 (aryl, preferably -SO 2 C 6 (aryl, such as -SO 2 -phenyl), -COC 6-10 (aryl, preferably -COC 6 (aryl, such as -CO-phenyl), a 4- to 6-membered saturated or unsaturated monocyclic heterocycle, a 4- to 6-membered saturated or unsaturated monocyclic ring, a 5- to 6-membered monocyclic heteroaryl ring, an 8- to 10-membered bicyclic heteroaryl ring, a spiro ring, a spiro heterocycle, a bridged ring or a bridged heterocycle, wherein R a0 , R b0 are each independently hydrogen or C 1-3 (alkyl).

[0182] Each of the various substituent groups described above herein may itself be substituted by the groups described herein.

[0183] When the 4- to 6-membered saturated monocyclic heterocycle described herein is substituted, the position of the substituent may be at their possible chemical positions, and representative substitution cases of exemplary monocyclic heterocycles are shown below:

[0184]

[0185]

[0186] wherein "Sub" represents the various substituent groups described herein; represents the connection to other atoms.

[0187] Pharmaceutical composition

[0188] Generally, the compounds of the present invention, or their pharmaceutically acceptable salts, or their solvates, or their stereoisomers, or prodrugs can be administered in a suitable dosage form in combination with one or more pharmaceutical carriers. These dosage forms are suitable for oral, rectal, topical, buccal, and other parenteral administrations (e.g., subcutaneous, intramuscular, intravenous, etc.). For example, dosage forms suitable for oral administration include capsules, tablets, granules, syrups, etc. The compounds of the present invention contained in these preparations can be solid powders or granules; solutions or suspensions in aqueous or non-aqueous liquids; water-in-oil or oil-in-water emulsions, etc. The above dosage forms can be prepared from the active compound and one or more carriers or excipients by common pharmaceutical methods. The above carriers need to be compatible with the active compound or other excipients. For solid preparations, common non-toxic carriers include but are not limited to mannitol, lactose, starch, magnesium stearate, cellulose, glucose, sucrose, etc. Carriers for liquid preparations include water, normal saline, glucose aqueous solution, ethylene glycol, polyethylene glycol, etc. The active compound can form a solution or suspension with the above carriers.

[0189] "Pharmaceutically acceptable carrier" refers to a non-toxic, inert, solid, semi-solid substance or liquid filling machine, diluent, encapsulating material or auxiliary preparation or any type of excipient, which is compatible with the patient, preferably a mammal, more preferably a human, and is suitable for delivering the active agent to the target site without terminating the activity of the agent.

[0190] "Active substance of the present invention" or "active compound of the present invention" refers to the compound of formula (I) of the present invention, or its pharmaceutically acceptable salt, or its solvate, or its stereoisomer, or its prodrug, which has a high selective inhibitory activity against CDK9.

[0191] The compositions of the present invention are formulated, quantified, and administered in a manner consistent with medical practice. The "therapeutically effective amount" of the compound administered is determined by factors such as the specific condition to be treated, the individual being treated, the cause of the condition, the target of the drug, and the mode of administration.

[0192] "Therapeutically effective amount" refers to the amount of the compound of the present invention that will cause a biological or medical response in an individual, such as reducing or inhibiting enzyme or protein activity, or improving symptoms, alleviating the condition, slowing down or delaying the disease process, or preventing the disease.

[0193] The therapeutically effective amount of the compound of the present invention, or its pharmaceutically acceptable salt, or its solvate, or its stereoisomer, or its prodrug contained in the pharmaceutical composition or the medicinal composition of the present invention is preferably 0.1 mg - 5 g / kg (body weight).

[0194] "Patient" refers to an animal, preferably a mammal, more preferably a human. The term "mammal" refers to warm-blooded vertebrate mammals, including, for example, cats, dogs, rabbits, bears, foxes, wolves, monkeys, deer, rats, pigs, and humans.

[0195] "Treatment" refers to alleviating, delaying progression, attenuating, preventing, or maintaining an existing disease or disorder (such as cancer). Treatment also includes curing one or more symptoms of a disease or disorder, preventing its development, or alleviating it to a certain extent.

[0196] The "pharmaceutically acceptable salts" include pharmaceutically acceptable acid addition salts and pharmaceutically acceptable base addition salts. Pharmaceutically acceptable acid addition salts refer to salts formed with inorganic acids or organic acids that can retain the biological effectiveness of the free base without other side effects. These salts can be prepared by methods known in the art.

[0197] "Pharmaceutically acceptable base addition salts" include, but are not limited to, salts of inorganic bases and salts of organic bases. These salts can be prepared by methods known in the art.

[0198] The "solvates" mentioned in the present invention refer to complexes formed by the compounds of the present invention with solvents. They either react in solvents or precipitate or crystallize out from solvents. For example, a complex formed with water is called a "hydrate". Solvates of the compounds of formula (I) are within the scope of the present invention.

[0199] When the compounds represented by formula (I) of the present invention contain one or more chiral centers, they can exist in different optically active forms. When the compound of formula (I) contains one chiral center, the compound comprises a pair of enantiomers. The two enantiomers of the compound and mixtures of the pair of enantiomers, such as racemic mixtures, are also within the scope of protection of the present invention. Enantiomers can be resolved by methods known in the art, such as crystallization and chiral chromatography. When the compound of formula (I) contains more than one chiral center, the compound comprises enantiomers and diastereomers. All enantiomers and diastereomers of the compound, as well as mixtures of enantiomers, mixtures of diastereomers, and mixtures of enantiomers and diastereomers are also within the scope of protection of the present invention. Enantiomers and diastereomers can be resolved by methods known in the art, such as crystallization and preparative chromatography.

[0200] The present invention includes prodrugs of the above-mentioned compounds. The prodrugs include known amino protecting groups and carboxyl protecting groups, and are hydrolyzed under physiological conditions or released via an enzymatic reaction to obtain the parent compound. Specific methods for preparing the prodrugs can be referred to (Saulnier, M.G.; Frennesson, D.B.; Deshpande, M.S.; Hansel, S.B and Vysa, D.M. Bioorg. Med. Chem Lett. 1994, 4, 1985-1990; and Greenwald, R.B.; Choe, Y.H.; Conover, C.D.; Shum, K.; Wu, D.; Royzen, M. J. Med. Chem. 2000, 43, 475.).

[0201] Preparation method

[0202] The present invention provides a method for preparing the compound of formula (I). The compound of formula (I) can be synthesized using standard synthetic techniques known to those skilled in the art or by combining methods known in the art with the methods described in the present invention. The solvents, temperatures and other reaction conditions given in the present invention can be changed according to the art. The reactions can be used sequentially to provide the compounds of the present invention, or they can be used to synthesize fragments, which are subsequently added by the methods described in the present invention and / or methods known in the art.

[0203] The compounds described in the present invention can be synthesized using methods similar to those described below or the exemplary methods described in the examples, or relevant published literature used by those skilled in the art, by using appropriate alternative starting materials. The starting materials for synthesizing the compounds described in the present invention can be synthesized or obtained from commercial sources. The compounds described in the present invention and other related compounds with different substituents can be synthesized using techniques and starting materials known to those skilled in the art. The general methods for preparing the compounds disclosed in the present invention can be derived from reactions known in the art, and the reactions can be modified by reagents and conditions considered appropriate by those skilled in the art to introduce various moieties into the molecules provided by the present invention.

[0204] Compared with the prior art, the main advantages of the present invention are as follows:

[0205] A series of structurally novel substituted bis(pyridin-2-yl)amine derivatives are provided, which have high selective inhibitory activity against CDK9, excellent in vivo pharmacokinetic activity and low in vivo toxicity, and thus can be used as drugs for treating and / or preventing diseases related to or mediated by CDK9 activity. Further, it can be used for oral administration.

[0206] The present invention will be further described below in conjunction with specific embodiments. It should be understood that these embodiments are only used to illustrate the present invention and not to limit the scope of the present invention. The experimental methods without specific conditions noted in the following embodiments are generally carried out under conventional conditions such as those described in Sambrook et al., Molecular Cloning: A Laboratory Manual (New York: Cold Spring Harbor Laboratory Press, 1989), or according to the conditions recommended by the manufacturer. Unless otherwise specified, percentages and parts are calculated by weight. Unless otherwise defined, the terms used herein have the same meaning as those familiar to those skilled in the art. In addition, any methods and materials similar or equivalent to those described can be applied to the present invention.

[0207] Reagents and Instruments

[0208] 1 HNMR: Bruker AVANCE-400 nuclear magnetic resonance instrument, with tetramethylsilane (TMS) as the internal standard.

[0209] LC-MS: Agilent 1290 HPLC System / 6130 / 6150 MS liquid chromatography-mass spectrometry instrument (manufacturer: Agilent), column Waters BEH / CHS, 50×2.1 mm, 1.7 μm.

[0210] Preparative high performance liquid chromatography (pre-HPLC): GX-281 (manufacturer: Gilson).

[0211] ISCO Combiflash-Rf75 or Rf200 type automatic column chromatograph is used, and Agela 4g, 12g, 20g, 40g, 80g, 120g disposable silica gel columns.

[0212] Known starting materials can be used or synthesized according to methods known in the art, or can be purchased from companies such as ABCR GmbH & Co. KG, Acros Organics, Aldrich Chemical Company, Accela ChemBio Inc, and Darui Chemicals.

[0213] In the examples, the progress of the reaction can be monitored by thin layer chromatography (TLC), and the compound purification can be carried out by column chromatography. The developing agent systems used in column chromatography or TLC can be selected from: dichloromethane and methanol system, n-hexane and ethyl acetate system, petroleum ether and ethyl acetate system, and acetone system, etc. The volume ratio of the solvents is adjusted according to the polarity of the compounds.

[0214] As used herein, DMF: N,N-dimethylformamide, DMSO: dimethyl sulfoxide, THF: tetrahydrofuran, DIEA: N,N-diisopropylethylamine, EA: ethyl acetate, PE: petroleum ether, BINAP: (2R,3S)-2,2'-bis(diphenylphosphino)-1,1'-binaphthyl, NBS: N-bromosuccinimide, NCS: N-chlorosuccinimide, Pd 2 (dba) 3 : tris(dibenzylideneacetone)dipalladium, Pd(dppf)Cl 2 : [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium, Pd(PPh 3 ) 4 : tetrakis(triphenylphosphine)palladium, PdCl 2 (CH 3 CN) 2： dichlorobis(acetonitrile)palladium, DPPA: diphenylphosphoryl azide, DBU: 1,8-diazabicyclo[5.4.0]undec-7-ene, TBAF: tetrabutylammonium fluoride, Na Ascorbate: sodium ascorbate, t-BuXPhosPd-G3: palladium(II) (2-di-tert-butylphosphino-2',4',6'-triisopropyl-1,1'-biphenyl)(2'-amino-1,1'-biphenyl-2-yl) methanesulfonate, X-PHOS: 2-dicyclohexylphosphino-2',4',6'-triisopropylbiphenyl, XantPhos: 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene.

[0215] As used herein, room temperature refers to about 20 - 30 °C.

[0216] Intermediate v1

[0217]

[0218] Compound 1-1 (9.6 g, 37.55 mmol), Pd 2 (dba) 3(687.74 mg, 751.03 μmol), X-PHOS (716.07 mg, 1.50 mmol) were dissolved in THF (40 mL). The mixture was purged with argon three times, and then lithium bis(trimethylsilyl)amide (12.57 g, 75.10 mmol, 1 M in THF) was slowly added. After the addition, the reaction mixture was stirred at 60 °C for 6 h. The reaction mixture was poured into 240 mL of 1 M hydrochloric acid, stirred for 30 minutes, adjusted to pH = 8 with saturated sodium bicarbonate, extracted with EA (150 mL × 2), the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue obtained was purified by silica gel column chromatography with an eluent of 0 - 40% EA in PE to obtain intermediate compound v1 (6.25 g). MS m / z (ESI): 237.0 [M+1].

[0219] Intermediate v2

[0220]

[0221] Compound 1-3 (1.15 g, 3.18 mol) was dissolved in THF (5 mL), and ammonia methanol solution (30 mL, 1 M) was added. The mixture was stirred at 60 °C for 16 h under a sealed tube. The reaction mixture was concentrated under reduced pressure, the residue was dissolved in 20 mL of DCM, adjusted to pH = 8 with saturated aqueous sodium bicarbonate, extracted with DCM (30 mL × 2), the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue obtained was purified by silica gel column chromatography with an eluent of (0 - 30% methanol in DCM) to obtain intermediate compound v2 (900 mg). MS m / z (ESI): 343.1 [M+1].

[0222] Example 1: Preparation of Compound Z-1

[0223]

[0224] Step 1: Under nitrogen protection, 2-chloro-5-fluoro-4-iodopyridine (20 g, 77.69 mmol), (4-fluoro-2-methoxyphenyl)boronic acid (17.16 g, 101.00 mmol), Pd(PPh 3 ) 4(8.98 g, 7.77 mmol), potassium carbonate (32.16 g, 233.07 mmol), 1,4-dioxane (400 mL) and water (100 mL) were heated to 100 °C and reacted for 16 h. The reaction solution was concentrated to dryness, ethyl acetate (50 mL) was added, and it was extracted with distilled water (100 mL × 2). The organic phase was washed with saturated brine, concentrated, and purified by column chromatography (PE:EA = 98%:2%) to obtain compound 1-1 (14.1 g). MS m / z (ESI): 256.0 [M+1].

[0225] Step 2: Under nitrogen protection, compound 1-1 (7 g, 27.38 mmol), (2-amino-4-pyridyl)methanol (3.57 g, 28.75 mmol), Pd 2 (dba) 3 (2.51 g, 2.74 mmol), X-PHOS (2.61 g, 5.48 mmol), cesium carbonate (17.85 g, 54.76 mmol) and 1,4-dioxane (150 mL) were placed in a reaction flask and heated at 120 °C for 16 h. The reaction solution was filtered, the filtrate was evaporated to dryness, water was added to the residue and it was extracted with ethyl acetate (15 mL × 2). The organic layer was dried over anhydrous sodium sulfate and then evaporated to dryness to obtain the crude product. The crude product was purified by column chromatography (PE:EA = 90%:10% - 30%:70%) to obtain compound 1-2 (6.59 g). MS m / z (ESI): 344.0 [M+1].

[0226] Step 3: Thionyl chloride (11.42 g, 95.97 mmol, 6.97 mL) was added dropwise to a solution of compound 1-2 (6.59 g, 19.19 mmol) and THF (100 mL) under an ice bath. After addition, it was heated to 50 °C and reacted for 1.5 h. After the reaction solution was evaporated to dryness, saturated sodium bicarbonate solution was added under an ice bath to adjust the pH to 9, and then it was extracted with EA (75 mL × 3). The extraction solution was filtered to obtain a white solid. The organic phase was separated, washed with saturated brine, dried over anhydrous sodium sulfate, and then evaporated to dryness to obtain compound 1-3 (6.1 g). MS m / z (ESI): 362.0 [M+1].

[0227] Step 4: Mix compound 1-3 (2 g, 5.53 mmol), thiourea (589.15 mg, 7.74 mmol), ethyl acrylate (774.86 mg, 7.74 mmol), sodium carbonate (820.41 mg, 7.74 mmol), polyethylene glycol 200 (60 mL), and water (3 mL), then heat the mixture to 56 °C and react for 16 hours. Add ice water to the reaction solution, extract with EA (30 mL * 2), wash the organic phase with saturated brine, dry with anhydrous sodium sulfate, and evaporate to dryness to obtain the crude product. The crude product is purified by column chromatography (PE:EA = 80%:20% - 75%:25%) to obtain compound 1-4 (1.25 g). MS m / z (ESI): 460.1 [M+1].

[0228] Step 5: Add m-chloroperbenzoic acid (552.27 mg, 2.72 mmol, 85% purity) to a solution of compound 1-4 (1.25 g, 2.72 mmol) and DCM (20 mL) at 0 °C, and react at -20 °C for 10 minutes. Add 30% sodium thiosulfate solution to the reaction solution at 0 °C to quench the reaction, then extract with EA (15 mL * 2), wash the organic phase with saturated brine and evaporate to dryness to obtain the crude product. The crude product is purified by column chromatography (PE:EA = 80%:20% - 0:100%) to obtain compound 1-5 (1.15 g). MS m / z (ESI): 476.1 [M+1].

[0229] Step 6: Mix compound 1-5 (1 g, 2.10 mmol), tert-butyl carbamate (369.54 mg, 3.15 mmol), magnesium oxide (339.04 mg, 8.41 mmol), dirhodium tetraacetate (278.86 mg, 630.91 μmol), iodobenzene diacetate (1.35 g, 4.21 mmol), and DCM (30 mL), then react at 45 °C for 3 hours. Add tert-butyl carbamate (369.54 mg, 3.15 mmol), magnesium oxide (339.04 mg, 8.41 mmol), dirhodium tetraacetate (278.86 mg, 630.91 μmol), and iodobenzene diacetate (1.35 g, 4.21 mmol) again. After adding, react at 45 °C for 3 hours. Then add tert-butyl carbamate (369.54 mg, 3.15 mmol), magnesium oxide (339.04 mg, 8.41 mmol), dirhodium tetraacetate (278.86 mg, 630.91 μmol), and iodobenzene diacetate (1.35 g, 4.21 mmol) again. After adding, react at 45 °C for 3 hours. Filter the reaction solution and evaporate to dryness to obtain the crude product. The crude product is purified by column chromatography (PE:EA = 100%:0 - 50%:50%) to obtain compound 1-6 (700 mg). MS m / z (ESI): 591.2 [M+1].

[0230] Step 7: Add trifluoroacetic acid (2 mL) to a solution of compound 1-6 (429.99 mg, 503.05 μmol) and DCM (6 mL) at 0 °C. After addition, react at room temperature for 1 hour. Evaporate the reaction solution to dryness to obtain compound 1-7 (220 mg). MS m / z (ESI): 491.2 [M+1].

[0231] Step 8: Mix compound 1-7 (220 mg, 448.50 μmol) and acetic acid (5 mL), heat to 75 °C and react for 8 hours. Evaporate the reaction solution to dryness, add the residue to DCM (15 mL), and wash with water (10 mL * 2). Purify by HPLC preparative liquid chromatography (preparative column: 21.2 X 250 mm C18 column; system: 10 mM NH 4 HCO 3 H 2 O; wavelength: 254 / 214 nm) to obtain compound Z-1 (26.89 mg). MS m / z (ESI): 445.0 [M+1]. 1 1H NMR (400 MHz, DMSO-d6) δ 9.91 (s, 1H), 8.19 (dd, J = 2.8, 1.6 Hz, 2H), 7.73 (d, J = 5.4 Hz, 1H), 7.66 (s, 1H), 7.35–7.30 (m, 1H), 7.07 (dd, J = 11.4, 2.4 Hz, 1H), 6.90 (m, 2H), 4.99 (q, J = 13.6 Hz, 2H), 4.06 (m, 1H), 3.77 (s, 3H), 3.54–3.45 (m, 1H), 2.88–2.79 (m, 1H), 2.53 (m, 1H).

[0232] Example 2: Preparation of compound Z-2

[0233]

[0234] Step 1: Mix compound 1-3 (100 mg, 276.24 μmol), N-(N-tert-butoxycarbonylaminocarbamoyl)carbamic acid tert-butyl ester (85.85 mg, 331.49 μmol), potassium carbonate (114.36 mg, 828.72 μmol) and acetonitrile (3 mL), heat to 50 °C and react for 16 hours. Add water to the reaction solution, extract with EA (10 mL * 2), wash the organic phase, wash with saturated brine, dry over anhydrous sodium sulfate, and perform column chromatography (PE:EA = 90%:10% to 75%:25%) to obtain compound 2-1 (146 mg). MS m / z (ESI): 585.3 [M+1].

[0235] Step 2: Mix compound 2-1 (146 mg, 249.74 μmol), trifluoroacetic acid (1.5 mL) and DCM (3 mL), and react at room temperature for 1 hour. After evaporating the reaction solution to dryness, it was sent to preparative liquid chromatography (preparative column: 21.2X250 mm C18 column; system: 10 mM NH 4 HCO 3 H 2 O; wavelength: 254 / 214 nm) for purification to obtain compound Z-2 (24.96 mg). MS m / z (ESI): 385.1 [M+1]. 1 HNMR (400 MHz, DMSO-d6) δ 9.82 (s, 1H), 8.18 (d, J = 1.2 Hz, 1H), 8.10 (d, J = 5.2 Hz, 1H), 7.96 (s, 2H), 7.76 (d, J = 5.6 Hz, 1H), 7.51 (s, 1H), 7.32 (dd, J = 8.4, 6.8 Hz, 1H), 7.07 (dd, J = 11.6, 2.4 Hz, 1H), 6.89 (td, J = 8.4, 2.4 Hz, 1H), 6.72 (d, J = 4.8 Hz, 1H), 4.35 (s, 2H), 3.78 (s, 1H), 3.77 (s, 3H).

[0236] Example 3: Preparation of compound Z-3

[0237]

[0238] Step 1: Heat compound 1-3 (1 g, 2.76 mmol), thiourea (252.49 mg, 3.32 mmol) and ethanol (15 mL) to 90 °C and react for 10 hours. Solids precipitated from the reaction solution. After evaporating the solvent to dryness, a crude yellow solid was obtained. The crude product was slurried with PE:EA = 1:1 (30 mL) and then filtered to obtain compound 3-1 (1.1 g). MS m / z (ESI): 402.0 [M+1].

[0239] Step 2: Mix compound 3-1 (1 g, 2.49 mmol), methyl 4-bromobutyrate (586.24 mg, 3.24 mmol), potassium carbonate (1.03 g, 7.47 mmol), acetonitrile (10 mL) and DMF (3 mL), and react at room temperature for 3 days. Filter the reaction solution. After evaporating the filtrate to dryness, add water, and then extract with EA (15 mL * 2). Combine the organic phases, wash with saturated brine, dry over anhydrous sodium sulfate, filter, and evaporate to dryness to obtain a crude product. The crude product was purified by column chromatography (PE:EA = 90%:10% - 75%:25%) to obtain compound 3-2 (724 mg). MS m / z (ESI): 460.2 [M+1].

[0240] Step 3-6: Refer to the preparation method in steps 5-8 of Example 1, with the difference that compound 3-2 is used to replace compound 1-4, to obtain compound Z-3 (3.23 mg). MS m / z (ESI): 459.1 [M+1]. 1 H NMR (400 MHz, DMSO-d6) δ 9.88 (s, 1H), 8.18 (d, J = 4.6 Hz, 2H), 7.73 (d, J = 5.6 Hz, 1H), 7.65 (s, 1H), 7.34–7.30 (m, 1H), 7.06 (dd, J = 11.4, 2.4 Hz, 1H), 6.91–6.86 (m, 2H), 4.77 (d, J = 15.2 Hz, 2H), 3.76 (s, 3H), 3.64–3.55 (m, 2H), 3.22–3.15 (m, 2H), 2.28–2.22 (m, 1H), 2.08–2.04 (m, 1H).

[0241] Example 4: Preparation of compound Z-4

[0242]

[0243] Step 1: Mix 2-chloro-4-(chloromethyl)pyridine (500 mg, 3.09 mmol), 3-sulfanylpropan-1-ol (341.30 mg, 3.70 mmol), potassium carbonate (639.78 mg, 4.63 mmol) and acetonitrile (20 mL), and heat the mixture at 50 °C for 3 hours. Filter the reaction solution and evaporate it to dryness to obtain compound 4-1 (670 mg). MS m / z (ESI): 218.0 [M+1].

[0244] Step 2: Add m-chloroperbenzoic acid (624.77 mg, 3.08 mmol, 85% purity) to a solution of compound 4-1 (670.00 mg, 3.08 mmol) and DCM (20 mL) at 0 °C, and react at 0 °C for 10 minutes. Filter the reaction solution and evaporate it to dryness to obtain compound 4-2 (650 mg). MS m / z (ESI): 234.0 [M+1].

[0245] Step 3: tert-Butyldimethylsilyl chloride (1.26 g, 8.34 mmol) was added to a solution of compound 4-2 (650 mg, 2.78 mmol), imidazole (1.14 g, 16.69 mmol), 4-dimethylaminopyridine (339.77 mg, 2.78 mmol) and DCM (30 mL), and the reaction was carried out at room temperature for 2 hours. The reaction mixture was added to ice water, extracted with DCM, the organic layer was washed with saturated brine, dried and evaporated to dryness to obtain a crude product. The crude product was purified by column chromatography (PE:EA = 90%:10% - 55%:45%) to obtain compound 4-3 (950 mg), a yellow oil. MS m / z (ESI): 348.0 [M+1].

[0246] Step 4: Compound 4-3 (600 mg, 1.72 mmol), tert-butyl carbamate (302.99 mg, 2.59 mmol), magnesium oxide (277.98 mg, 6.90 mmol), dirhodium tetraacetate (228.64 mg, 517.28 μmol), iodobenzene diacetate (1.11 g, 3.45 mmol) and DCM (20 mL) were mixed and reacted at 45 °C for 8 hours. tert-Butyl carbamate (302.99 mg, 2.59 mmol), magnesium oxide (277.98 mg, 6.90 mmol), dirhodium tetraacetate (228.64 mg, 517.28 μmol), iodobenzene diacetate (1.11 g, 3.45 mmol) were added, and after addition, the reaction was carried out at 45 °C for 4 hours. The reaction mixture was filtered and evaporated to dryness to obtain a crude product. The crude product was purified by column chromatography (PE:EA = 100%:0% - 60%:40%) to obtain compound 4-4 (664 mg). MS m / z (ESI): 463.1 [M+1].

[0247] Step 5: Compound 4-4 (576 mg, 1.28 mmol), ammonium fluoride (950.09 mg, 25.65 mmol) and methanol (20 mL) were mixed and reacted at 50 °C for 2 hours. The reaction mixture was filtered, and the filtrate was evaporated to dryness to obtain a crude product. The crude product was slurried with DCM:methanol = 10:1 and filtered to remove inorganic salts, and the filtrate was evaporated to dryness to obtain compound 4-5 (412 mg). MS m / z (ESI): 349.0 [M+1].

[0248] Step 6: Trifluoroacetic acid (3 mL) was added to a solution of compound 4-5 (440 mg, 1.26 mmol) and DCM (9 mL) at 0 °C, and the reaction was carried out at room temperature for 1 hour. The reaction mixture was evaporated to dryness to obtain a crude product. The crude product was purified by reverse-phase column chromatography (aqueous ammonium bicarbonate solution / acetonitrile) to obtain compound 4-6 (310 mg). MS m / z (ESI): 249.0 [M+1].

[0249] Step 7: Methanesulfonyl chloride (87.50 mg, 763.88 μmol) was added to a solution of compound 4-6 (200 mg, 804.09 μmol), triethylamine (162.73 mg, 1.61 mmol, 224.30 μL) and DCM (10 mL) at 0 °C. After addition, the reaction was carried out at 0 °C for 2 hours. The reaction mixture was quenched with a small amount of ice water at 0 °C, and the solution was evaporated to dryness to obtain a crude product. The crude product was purified by column chromatography (reverse phase, aqueous ammonium bicarbonate / acetonitrile) to obtain compound 4-7 (138 mg). MS m / z (ESI): 327.0 [M+1].

[0250] Step 8: Compound 4-7 (138 mg, 422.25 μmol) and ammonia water (5 mL) were heated to 80 °C and reacted for 2 hours. The reaction mixture was evaporated to dryness to obtain compound 4-8 (95 mg). MS m / z (ESI): 231.0 [M+1].

[0251] Step 9: Under nitrogen protection, compound 4-8 (45 mg, 195.05 μmol), intermediate v1 (32.25 mg, 136.53 μmol), Pd2(dba)3 (17.87 mg, 19.50 μmol), XantPhos (22.55 mg, 39.01 μmol), potassium carbonate (53.91 mg, 390.09 μmol) and 1,4-dioxane (5 mL) were heated to 130 °C and reacted by microwave for 1 hour. The reaction mixture was filtered and the filtrate was evaporated to dryness to obtain a crude product. The crude product was prepared by preparative HPLC (preparation column: 21.2X250 mm C18 column; system: 10 mM NH 4 HCO 3 H 2 O; wavelength: 254 / 214 nm) to obtain compound Z-4 (1.16 mg). MS m / z (ESI): 431.1 [M+1]. 11H NMR (400 MHz, DMSO-d6) δ 9.78 (s, 1H), 8.18 (d, J = 1.4 Hz, 1H), 8.13 (d, J = 5.2 Hz, 1H), 7.75 (d, J = 5.4 Hz, 1H), 7.59 (s, 1H), 7.33–7.29 (m, 1H), 7.06 (d, J = 9.0 Hz, 1H), 6.92–6.89 (m, 1H), 6.87 (dd, J = 8.4, 2.4 Hz, 1H), 4.57 (d, J = 13.6 Hz, 1H), 4.45 (d, J = 13.6 Hz, 1H), 3.76 (s, 3H), 3.60–3.55 (m, 1H), 3.38 (m, 1H), 3.15 (m, 1H), 2.98–2.92 (m, 1H), 2.02–1.96 (m, 1H), 1.87–1.81 (m, 1H).

[0252] Example 4-1: Preparation of Enantiomers of Compound Z-4

[0253]

[0254] Compound Z-4 was separated by a chiral HPLC column (Waters SFC-150; column: OD-3 4.6*100 mm 3um solvent: methanol (0.2% NH 3 (7M in methanol)); flow rate: 3.0 ml / min;), and enantiomers with retention times of 2.284 min and 2.621 min were obtained respectively.

[0255] Example 5: Preparation of Compound Z-5

[0256]

[0257] Step 1: Compound 1-3 (200 mg, 552.83 μmol), 7M ammonia methanol solution (10 mL), and THF (3 mL) were heated to 50 °C and reacted for 4 hours, and then heated to 60 °C and reacted for 16 hours. The reaction solution was evaporated to dryness to obtain a crude product. The crude product was purified by column chromatography (PE: EA = 80%: 20% ~ 0:100%--DCM: methanol = 95%: 5%) to obtain Compound 5-1 (90 mg). MS m / z (ESI): 343.1 [M+1].

[0258] Step 2: Heat compound 5-1 (90 mg, 262.89 μmol), 2-methylthio-4,5-dihydro-1H-imidazole (64.17 mg, 262.89 μmol, hydroiodide), and pyridine (5 mL) to 125 °C and react for 2 hours. Evaporate the reaction solution to dryness to obtain the crude product. The crude product was purified by preparative HPLC (preparative column: 21.2 X 250 mm C18 column; system: 10 mM NH 4 HCO 3 H 2 O; wavelength: 254 / 214 nm) to obtain compound Z-5 (9.51 mg). MS m / z (ESI): 411.2 [M+1]. 1 1H NMR (400 MHz, DMSO-d6) δ 9.71 (s, 1H), 8.17 (s, 1H), 8.07 (d, J = 5.2 Hz, 1H), 7.77 (d, J = 5.4 Hz, 1H), 7.46 (s, 1H), 7.34–7.28 (m, 1H), 7.06 (d, J = 9.0 Hz, 1H), 6.88 (s, 1H), 6.73 (s, 1H), 4.29 (s, 2H), 3.76 (s, 3H), 3.43 (s, 4H).

[0259] Example 6: Preparation of compound Z-6

[0260]

[0261] Step 1: Heat compound 1-3 (200 mg, 552.83 μmol) and methylamine / THF (5 mL) to 60 °C in a sealed tube and react for 16 hours. Evaporate the reaction solution to dryness, then triturate with DCM, filter off the solid salt, and evaporate the filtrate to dryness to obtain compound 6-1 (190 mg). MS m / z (ESI): 357.1 [M+1].

[0262] Step 2: Heat compound 6-1 (50 mg, 140.30 μmol), 1H-pyrazole-1-carboximidamide hydrochloride (61.70 mg, 420.91 μmol, HCl), DIEA (72.53 mg, 561.22 μmol, 97.75 μL), and DMSO (5 mL) to 130 °C and react for 16 hr. The reaction solution was purified by preparative HPLC (preparative column: 21.2 X 250 mm C18 column; system: 10 mMNH 4 HCO 3 H 2 O; wavelength: 254 / 214 nm) to obtain compound Z-6 (13.04 mg). MS m / z (ESI): 399.2 [M+1]. 1HNMR (400 MHz, DMSO-d6) δ 9.81 (s, 1H), 8.41 (s, 1H), 8.29 (s, 2H), 8.19 (d, J = 1.6 Hz, 1H), 8.13 (d, J = 5.2 Hz, 1H), 7.76 (d, J = 5.4 Hz, 1H), 7.45 (s, 1H), 7.31 (dd, J = 8.4, 6.8 Hz, 1H), 7.07 (dd, J = 11.4, 2.4 Hz, 1H), 6.89 (td, J = 8.4, 2.4 Hz, 1H), 6.62 (dd, J = 5.2, 1.4 Hz, 1H), 4.53 (s, 2H), 3.76 (s, 3H), 2.94 (s, 3H).

[0263] Example 7: Preparation of Compound Z-7

[0264]

[0265] Step 1: Ethyl N-(thiomethylene)carbamate (3375 mg, 2.86 mmol), 2-(tert-butoxycarbonylamino)acetic acid (500.89 mg, 2.86 mmol), pyridine (452.33 mg, 5.72 mmol, 460.67 μL) and acetonitrile (15 mL) were reacted at room temperature for 16 hours. The reaction solution was evaporated to dryness to obtain a crude product. The crude product was purified by column chromatography (PE:EA = 85%:15%) to obtain Compound 7-1 (580 mg). MS m / z (ESI): 161.0 [M-56+1].

[0266] Step 2: Compound 7-1 (220 mg, 1.02 mmol), potassium carbonate (281.20 mg, 2.03 mmol) and acetonitrile (10 mL) were mixed, then methyl iodide (144.40 mg, 1.02 mmol) was added, and after addition, the mixture was reacted at room temperature for 16 hours. The reaction solution was filtered and evaporated to dryness to obtain a crude product, and the crude product was slurried with distilled water and then filtered and evaporated to dryness to obtain Compound 7-2 (280 mg). MS m / z (ESI): 175.0 [M-56+1].

[0267] Step 3: Intermediate v2 (50 mg, 146.05 μmol), Compound 7-2 (50.45 mg, 219.08 μmol) and ethanol (5 mL) were heated at 60 °C for 48 hours. The reaction solution was evaporated to dryness to obtain Compound 7-3 (76 mg). MS m / z (ESI): 525.2 [M+1].

[0268] Step 4: Add trifluoroacetic acid (0.8 mL) to a solution of compound 7-3 (71 mg, 135.36 μmol) and DCM (2 mL), and react at room temperature for 1 hour. Evaporate the reaction solution to dryness to obtain the crude product. The crude product was prepared by preparative HPLC (preparation column: 21.2 X 250 mm C18 column; system: 10 mM NH 4 HCO 3 H 2 O; wavelength: 254 / 214 nm) to obtain compound Z-7 (5.93 mg). MS m / z (ESI): 425.0 [M+1]. 1 1H NMR (400 MHz, DMSO-d6) δ 9.74 (s, 1H), 8.43 (s, 1H), 8.16 (d, J = 1.2 Hz, 1H), 8.08 (d, J = 5.2 Hz, 1H), 7.94 (s, 1H), 7.78 (d, J = 5.6 Hz, 1H), 7.45 (s, 1H), 7.31 (dd, J = 8.4, 6.8 Hz, 1H), 7.06 (dd, J = 11.4, 2.4 Hz, 1H), 6.88 (td, J = 8.4, 2.4 Hz, 1H), 6.72 (dd, J = 5.2, 1.2 Hz, 1H), 4.36 (s, 2H), 3.76 (s, 3H), 3.65 (s, 2H).

[0269] Example 8: Preparation of compound Z-8

[0270]

[0271] Heat compound 1-3 (200 mg, 552.83 μmol), 1H-benzoimidazol-2-amine (73.61 mg, 552.83 μmol), potassium hydroxide (62.03 mg, 1.11 mmol) and acetone (5 mL) to 60 °C and react for 3 hours. After evaporating the reaction solution to dryness, add water (5 mL), extract with EA (5 mL * 2), combine the organic layers, wash with saturated brine and evaporate to dryness to obtain the crude product. The crude product was purified by column chromatography (DCM: methanol = 95%: 5%) to obtain compound Z-8 (113 mg). MS m / z (ESI): 459.1 [M+1]. 11H NMR (400 MHz, DMSO-d6) δ 9.73 (s, 1H), 8.08 (d, J = 1.2 Hz, 1H), 8.04 (d, J = 5.2 Hz, 1H), 7.70 (d, J = 5.4 Hz, 1H), 7.39 (s, 1H), 7.28 (dd, J = 8.4, 6.8 Hz, 1H), 7.13 (d, J = 7.6 Hz, 1H), 7.05 (dd, J = 11.4, 2.4 Hz, 1H), 6.98 (d, J = 7.6 Hz, 1H), 6.92 (td, J = 7.4, 1.0 Hz, 1H), 6.86 (td, J = 8.4, 4.2 Hz, 1H), 6.80 (td, J = 7.6, 0.8 Hz, 1H), 6.48 (m, 2H), 5.22 (s, 2H), 3.75 (s, 3H).

[0272] Example 9: Preparation of Compound Z-9

[0273]

[0274] Compound 1-3 (100 mg, 276.42 μmol), N-methyl-1H-benzimidazol-2-amine (40.68 mg, 276.42 μmol), potassium hydroxide (31.02 mg, 552.83 μmol) and acetone (15 mL) were heated to 60 °C and reacted for 2 hours. The reaction solution was evaporated to dryness, the residue was added with water (5 mL) and extracted with EA (5 mL * 3). The combined organic phases were washed with saturated brine and then evaporated to dryness to obtain the crude product. The crude product was purified by column chromatography (DCM: methanol = 95%: 5%) to obtain 60 mg of a relatively pure product, and then further purified by preparative HPLC (preparative column: 21.2X250 mm C18 column; system: 10 mM NH 4 HCO 3 H 2 O; wavelength: 254 / 214 nm) to obtain Compound Z-9 (13.33 mg). MS m / z (ESI): 473.1 [M + 1]. 11H NMR (400 MHz, DMSO-d6) δ 9.74 (s, 1H), 8.08 (d, J = 1.6 Hz, 1H), 8.04 (d, J = 5.2 Hz, 1H), 7.70 (d, J = 5.4 Hz, 1H), 7.35 (s, 1H), 7.31–7.25 (m, 1H), 7.20 (d, J = 7.2 Hz, 1H), 7.05 (dd, J = 11.4, 2.4 Hz, 1H), 7.00 (d, J = 7.6 Hz, 1H), 6.92 (td, J = 7.6, 1.2 Hz, 1H), 6.87 (td, J = 8.4, 2.4 Hz, 1H), 6.82 (td, J = 7.6, 1.2 Hz, 1H), 6.73 (d, J = 4.8 Hz, 1H), 6.47 (dd, J = 5.2, 1.2 Hz, 1H), 5.20 (s, 2H), 3.75 (s, 3H), 2.89 (d, J = 4.8 Hz, 3H).

[0275] Example 10: Preparation of Compound Z-10

[0276]

[0277] Step 1: Under nitrogen protection, cool 2-chloro-5-methoxypyridine (500 mg, 3.48 mmol) and THF (15 mL) to -78 °C, add lithium diisopropylamide (2 M, 3.48 mL), react at -78 °C for 1 hour after addition, then add DMF (509.11 mg, 6.97 mmol, 539.31 μL), and react at -78 °C for 1 hour after addition. Quench the reaction mixture with saturated ammonium chloride solution and extract (10 mL * 2). Wash the organic layer with saturated brine and evaporate to dryness to obtain Compound 10-1 (530 mg). MS m / z (ESI): 190.0 [M + 18 + 1].

[0278] Step 2: Add sodium borohydride (229.31 mg, 6.06 mmol) to a solution of Compound 10-1 (520 mg, 3.03 mmol) and methanol (1 mL) at 0 °C and react at 0 °C for 10 minutes. Quench the reaction with 1 N hydrochloric acid at 0 °C, evaporate the reaction mixture to dryness and extract with DCM (10 mL * 2). Wash the organic phase with saturated brine, dry over anhydrous sodium sulfate and evaporate to dryness to obtain the crude product. Purify the crude product by column chromatography (PE:EA = 70%:30% - 50%:50%) to obtain Compound 10-2 (370 mg). MS m / z (ESI): 174.0 [M + 1].

[0279] Step 3-9: Referring to the preparation method in Step 2-8 of Example 1, except that compound 10-2 is used to replace compound 1-1, and intermediate v1 is used to replace (2-amino-4-pyridyl) methanol, to obtain compound Z-10 (1.10 mg). MS m / z (ESI): 475.1 [M+1]. 1 H NMR (400 MHz, DMSO-d6) δ 9.69 (s, 1H), 8.14 (s, 1H), 8.04 (s, 1H), 7.67 (s, 1H), 7.62 (d, J = 5.2 Hz, 1H), 7.31 (dd, J = 8.4, 6.8 Hz, 1H), 7.06 (dd, J = 11.4, 2.4 Hz, 1H), 6.90–6.86 (m, 1H), 4.96 (s, 2H), 4.18–4.11 (m, 1H), 3.84 (s, 3H), 3.76 (s, 3H), 3.58–3.52 (m, 1H), 2.88–2.82 (m, 1H), 2.63–2.57 (m, 1H).

[0280] Example 11: Preparation of compound Z-11

[0281]

[0282] Compound Z-8 (60 mg, 130.87 μmol), acetic anhydride (26.72 mg, 261.74 μmol) and THF (15 mL) were mixed and reacted at room temperature for 16 hours. The reaction solution was poured into ice water, extracted with EA (10 mL * 2), the organic layer was washed with saturated brine, dried over anhydrous sodium sulfate and evaporated to dryness to obtain a crude product. The crude product was purified by column chromatography (DCM: methanol = 95%: 5%) to obtain 40 mg of a relatively pure product, and then compound Z-11 (20.82 mg) was prepared by preparative HPLC liquid chromatography (preparative column: 21.2X250 mm C18 column; system: 10 mM NH4HCO3 H2O; wavelength: 254 / 214 nm). MS m / z (ESI): 501.1 [M+1]. 11H NMR (400 MHz, DMSO-d6) δ 10.53 (s, 1H), 9.73–9.69 (m, 1H), 8.05 (s, 2H), 7.69 (dd, J = 15.6, 6.2 Hz, 1H), 7.57–7.47 (m, 1H), 7.37 (d, J = 20.6 Hz, 2H), 7.28 (dd, J = 8.4, 6.8 Hz, 1H), 7.19–7.13 (m, 2H), 7.05 (dd, J = 11.4, 2.4 Hz, 1H), 6.87 (td, J = 8.4, 2.4 Hz, 1H), 6.74 and 6.45 (m, 1H), 5.30 (d, J = 13.2 Hz, 2H), 3.75 (s, 3H), 2.05 (d, J = 21.2 Hz, 3H).

[0283] Example 12: Preparation of Compound Z-12

[0284]

[0285] Dissolve intermediate v2 (150 mg, 438.16 μmol) in pyridine (4 mL), and add 1-cyano-2,3-dimethyl-isothiourea (56.60 mg, 438.16 μmol). React at 145 °C under microwave for 3.5 hours. Add 30 mL of water to the reaction solution, extract with EA (30 mL × 2), combine the organic phases, wash with saturated brine, dry over anhydrous sodium sulfate, filter, concentrate the filtrate under reduced pressure, and purify the residue by preparative liquid chromatography (preparative column: 21.2X250 mm C18 column; system: 10 mM NH 4 HCO 3 H 2 O; wavelength: 254 / 214 nm; gradient: 5% - 95% acetonitrile change) to obtain product Z-12 (19.9 mg). MS m / z (ESI): 424.1 [M+1]. 1 1H NMR (400 MHz, DMSO-d6) δ 9.77 (s, 1H), 8.21 (s, 1H), 8.10 (d, J = 5.2 Hz, 1H), 7.84 (d, J = 5.3 Hz, 1H), 7.57 (t, J = 5.9 Hz, 1H), 7.48 (s, 1H), 7.35 (t, J = 7.6 Hz, 1H), 7.15 (d, J = 4.6 Hz, 1H), 7.10 (dd, J = 11.5, 1.7 Hz, 1H), 6.92 (dd, J = 9.3, 7.3 Hz, 1H), 6.72 (d, J = 5.1 Hz, 1H), 4.30 (d, J = 5.9 Hz, 2H), 3.80 (s, 3H), 2.76 (d, J = 4.4 Hz, 3H).

[0286] Example 13: Preparation of Compound Z-13

[0287]

[0288] Step 1: In a 15 mL sealed tube, intermediate v2 (330 mg, 963.95 μmol) and 2-methylthio-4,5-dihydro-1H-imidazole (258.83 mg, 1.06 mmol, HI) were dissolved in pyridine (8 mL). The reaction was stirred at 120 °C for 3 hours. After completion of the reaction, it was cooled to room temperature and the solvent was removed by rotary evaporation under reduced pressure to obtain a crude product. The crude product was purified by silica gel column chromatography (DCM: methanol = 10:90) to obtain compound 13-1 (330 mg). MS m / z (ESI): 411.4 [M+1].

[0289] Step 2: Under argon protection, compound 13-1 (100 mg, 243.65 μmol) was dissolved in anhydrous DMF (3 mL), and then cesium carbonate (158.77 mg, 487.31 μmol) was added thereto. The mixture was stirred at room temperature for half an hour. Subsequently, 1,2-dibromoethane (68.66 mg, 365.48 μmol) was added dropwise to the reaction, and the reaction was stirred at room temperature for another 3 hours. After completion of the reaction, it was cooled to room temperature, filtered, and the solvent was removed by rotary evaporation to obtain a crude product. The crude product was purified by preparative liquid chromatography to obtain product Z-13 (1.54 mg). MS m / z (ESI): 437.5 [M+1]. 1 H NMR (400 MHz, CD3OD) δ 8.53 (s, 1H), 8.16 (d, J = 5.2 Hz, 1H), 8.09 (s, 1H), 7.62 (s, 1H), 7.55 (d, J = 5.2 Hz, 1H), 7.32–7.28 (m, 1H), 6.92 (dd, J = 11.2, 2.4 Hz, 1H), 6.82–6.77 (m, 2H), 4.50 (s, 2H), 4.07 (t, J = 8.0 Hz, 2H), 3.95 (t, J = 8.0 Hz, 2H), 3.81 (s, 3H), 3.50 (m, 4H).

[0290] Example 14: Preparation of Compound Z-14

[0291]

[0292] Step 1: Dissolve intermediate v2 (360 mg, 1.05 mmol) in anhydrous THF (10 mL), cool to 0 °C, add pyridine (124.77 mg, 1.58 mmol, 127.07 μL) thereto, and then slowly add o-phenylmethyl chloromethane sulfate (217.84 mg, 1.26 mmol) dropwise thereto. The reaction is stirred at 0 °C for 30 minutes. After completion of the reaction, dilute with water, extract with EA, wash the organic phase with saturated brine, dry over anhydrous sodium sulfate, and rotary evaporate the solvent under reduced pressure to obtain a crude product. The crude product is purified by silica gel column chromatography (PE:EA = 40:60) to obtain compound 14-1 (400 mg). MS m / z (ESI): 479.5 [M+1].

[0293] Step 2: Dissolve compound 14-1 (400 mg, 835.92 μmol) in anhydrous DMF (5 mL), and then add [3-(aminomethyl)oxetan-3-yl]methanol (195.85 mg, 1.67 mmol) and triethylamine (169.17 mg, 1.67 mmol) thereto. The reaction is stirred at 60 °C for 5 hours. After completion of the reaction, rotary evaporate the solvent under reduced pressure to obtain compound 14-2 (360 mg). MS m / z (ESI): 502.5 [M+1].

[0294] Step 3: In a 15 mL sealed tube, dissolve compound 14-2 (220 mg, 438.64 μmol) in acetone (5 mL), and then successively add methyl iodide (249.04 mg, 1.75 mmol) and DIEA (113.38 mg, 877.28 μmol, 152.80 μL) thereto. The reaction is stirred at 55 °C for 3 hours. After completion of the reaction, rotary evaporate the solvent under reduced pressure to obtain compound 14-3 (200 mg). MS m / z (ESI): 516.6 [M+1].

[0295] Step 4: Dissolve compound 14-3 (200 mg, 387.9 μmol) in anhydrous THF (10 mL), and then add sodium hydroxide (31 mg, 775.8 μmol) thereto. The reaction is stirred at room temperature for 5 hours. After completion of the reaction, filter off the inorganic salts, dilute with water, extract with EA, wash the organic phase with saturated brine, dry over anhydrous sodium sulfate, and rotary evaporate the solvent under reduced pressure to obtain a crude product. The crude product is purified by preparative liquid chromatography to obtain product Z-14 (16.05 mg). MS m / z (ESI): 468.5 [M+1]. 1 H NMR (400 MHz, DMSO-d 6)δ9.68(s,1H),8.16(d,J=6.0Hz,2H),8.03(d,J=4.8Hz,1H),7.78(d,J=5.6Hz,1H),7.39(s,1H),7.33–7.28(m,1H),7.06(dd,J=11.2,2.4Hz,1H),6.88(td,J=8.4,2.4Hz,1H),6.68(d,J=5.2Hz,1H),4.36(d,J=5.6Hz,2H),4.29(m,4H),4.10(s,2H),3.76(s,3H),3.38(s,2H).

[0296] Example 15: Preparation of Compound Z-15

[0297]

[0298] Dissolve compound Z-3 (130 mg) in THF (10 mL), add a solution of borane (1 M, 1.42 mL) in THF, and react at room temperature for 3 h. While the reaction mixture is under ice bath, add methanol dropwise until no more bubbles are generated. Rotate the reaction mixture to dryness, dissolve the residue in 4 mL of DCM and 1 mL of trifluoroacetic acid, and stir for 2 h. Adjust the pH of the residue to 8 with sodium bicarbonate, extract with DCM and rotate to dryness. Purify by preparative liquid chromatography (preparative column: 21.2X250 mm C18 column; system: 10 mM NH 4 HCO 3 H 2 O; wavelength: 254 / 214 nm; gradient: 5%-95% acetonitrile change) to obtain compound Z-15 (34.2 mg), MS m / z (ESI): 445.1 [M+1], 1H NMR (400 MHz, DMSO-d6) δ9.80(s,1H),8.17(s,1H),8.11(d,J=4.0Hz,1H),7.75(d,J=8.0,1H),7.56(s,1H),7.05(dd,J=16.0,4.0Hz 1H),6.90–6.84(m,3H),4.36(dd,J=48.0,13.6Hz,2H),3.75(s,3H),3.23–3.05(m,3H),2.75–2.69(m,1H),2.09-2.05(m,2H),1.48-1.44(m,2H).

[0299] Example 16: Preparation of Compound Z-16

[0300]

[0301] Refer to the preparation method of Reference Example 3, with the difference that methyl 5-bromovalerate is used to replace methyl 4-bromobutyrate to obtain compound Z-16. MS m / z(ESI): 473.1[M+1]; 1H NMR(400MHz, DMSO-d6)δ9.88(s, 1H), 8.17(d, J = 4.0Hz, 2H), 7.73(d, J = 4.0Hz, 1H), 7.65(s, 1H), 7.31(dd, J = 8.4, 6.8Hz, 1H), 7.06(dd, J = 12.0, 4.0Hz, 1H), 6.91 - 6.88(m, 2H), 4.71(dd, J = 26.0, 13.6Hz, 2H), 3.76(s, 3H), 3.74 - 3.69(m, 1H), 3.53–3.44(m, 1H), 2.62 - 2.50(m, 1H), 2.48 - 2.36(m, 1H), 2.09–2.03(m, 2H), 1.9 - 1.83(m, 1H), 1.59 - 1.49(m, 1H).

[0302] Example 17: Preparation of Compound Z-17

[0303]

[0304] Refer to the preparation method of Reference Example 15, with the difference that Z-16 is used to replace compound Z-3 to obtain compound Z-17. MS m / z(ESI): 459.1[M+1]; 1H NMR(400MHz, DMSO-d6)δ9.78(s, 1H), 8.18(s, 1H), 8.12(d, J = 8.0Hz, 1H), 7.77(d, J = 8.0Hz, 1H), 7.57(s, 1H), 7.32(dd, J = 12.0, 4.0Hz, 1H), 7.06(dd, J = 16.0, 4.0Hz, 1H), 6.91–6.86(m, 2H), 4.31(dd, J = 48.0, 12.0Hz, 2H), 3.76(s, 3H), 3.50 - 3.45(m, 1H), 3.21 - 3.07(m, 2H), 2.98 - 2.94(m, 1H), 1.69–1.51(m, 6H).

[0305] Example 18: Preparation of Compound Z-18

[0306]

[0307] Compound 1-3 (80 mg, 221.13 μmol), 1H-imidazol-2-amine (55.12 mg, 663.40 μmol), DIEA (142.90 mg, 1.11 mmol, 192.59 μL), potassium iodide (11.01 mg, 66.34 μmol), and DMF (5 mL) were stirred at room temperature for 16 h. The reaction mixture was poured into ice water and extracted with EA (10 mL×2). The organic layer was washed with water (5 mL×3) and saturated brine (5 mL), and concentrated to obtain the crude product. The crude product was purified by preparative liquid chromatography (preparative column: 21.2X250 mm C18 column; system: 10 mM NH4HCO3 H2O; wavelength: 254 / 214 nm) to give compound Z-18 (33.16 mg). MS m / z (ESI): 409.1 [M+1]; 1H NMR (400 MHz, DMSO-d6) δ 9.74 (s, 1H), 8.14 (d, J = 1.6 Hz, 1H), 8.06 (d, J = 5.2 Hz, 1H), 7.72 (d, J = 5.4 Hz, 1H), 7.34 (s, 1H), 7.29 (dd, J = 8.4, 6.8 Hz, 1H), 7.05 (dd, J = 11.6, 2.4 Hz, 1H), 6.87 (td, J = 8.4, 2.4 Hz, 1H), 6.54 (d, J = 1.2 Hz, 1H), 6.46 (dd, J = 5.2, 1.2 Hz, 1H), 6.39 (d, J = 1.6 Hz, 1H), 5.35 (s, 2H), 4.91 (s, 2H), 3.75 (s, 3H).

[0308] Example 19: Preparation of Compound Z-19

[0309]

[0310] Under nitrogen protection, compound 1-3 (50 mg, 138.21 μmol) and 1-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyrazole (57.51 mg, 276.42 μmol) were dissolved in water (2 mL) and ethylene glycol dimethyl ether (5 mL) and placed in a 15 mL microwave tube. Then Pd(dppf)Cl 2(15.97 mg, 13.82 μmol), sodium carbonate (36.62 mg, 345.52 μmol) were added thereto in sequence, and the mixture was evacuated by bubbling with N2 for 1 minute. The reaction was stirred at 100 °C for 4 hours. After completion of the reaction, it was extracted with EA, the organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and the solvent was rotary evaporated under reduced pressure to obtain the crude product. The crude product was purified by preparative chromatography to obtain compound Z-19 (1.85 mg). MS m / z (ESI): 408.4, [M+1]; 1H NMR (400 MHz, DMSO-d6) δ 9.68 (s, 1H), 8.14 (d, J = 1.6 Hz, 1H), 8.05 (d, J = 5.2 Hz, 1H), 7.76 (d, J = 5.2 Hz, 1H), 7.36 (d, J = 1.6 Hz, 1H), 7.31–7.27 (m, 2H), 7.05 (dd, J = 11.2, 2.4 Hz, 1H), 6.87 (td, J = 8.4, 2.4 Hz, 1H), 6.62 (d, J = 5.2 Hz, 1H), 5.99 (d, J = 1.6 Hz, 1H), 3.97 (s, 2H), 3.75 (s, 3H), 3.66 (s, 3H).

[0311] Example 20: Preparation of Compound Z-20

[0312]

[0313] Step 1: Under nitrogen protection, the compound (2 g, 5.53 mmol) and triisopropyl(prop-1-ynyl)silane (4.34 g, 22.11 mmol) were dissolved in acetonitrile (30 mL), and then PdCl 2 (CH 3 CN) 2 (430.26 mg, 1.66 mmol), XantPhos (789.45 mg, 1.66 mmol), cesium carbonate (3.60 g, 11.06 mmol) were added thereto in sequence. After the reaction was purged with argon 3 times at room temperature, it was heated to 75 °C and reacted for 2 hours. After completion of the reaction, it was cooled to room temperature, the reaction solution was extracted with EA, the organic phases were combined, dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (PE:EA = 70:30) to obtain compound 20-1 (1.4 g). MS m / z (ESI): 508.7 [M+1]

[0314] Step 2: Dissolve compound 20-2 (1.4 g, 2.76 mmol) in water (5 mL) and methanol (100 mL), then add ammonium fluoride (5.10 g, 137.88 mmol) thereto. The reaction was stirred under reflux for 4 hours. After completion of the reaction, the reaction solution was concentrated under reduced pressure, diluted with water, extracted with EA, the organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and the solvent was rotary evaporated under reduced pressure to obtain compound 20-2 (750 mg). MS m / z (ESI): 352.4 [M+1].

[0315] Step 3: Under N 2 protection, dissolve compound 20-2 (100 mg, 284.62 μmol) and 2-(6-azido-2-pyridyl)propan-2-ol (60.86 mg, 341.54 μmol) in THF (4 mL), tert-butanol (8 mL), and water (4 mL), then add copper sulfate pentahydrate (56.85 mg, 227.69 μmol), sodium ascorbate (56.39 mg, 284.62 μmol), and 0.5 mL of triethylamine thereto in sequence. After the reaction was purged with argon 3 times at room temperature, it was heated to 70 °C and reacted for 6 hours. After completion of the reaction, it was quenched with water, filtered through diatomaceous earth, and the solvent was rotary evaporated under reduced pressure to obtain a crude product. The crude product was purified by preparative chromatography to obtain compound Z-20 (9.88 mg). MS m / z (ESI): 544.6, [M+1]; 1 H NMR (400 MHz, DMSO-d 6 ) δ 9.66 (s, 1H), 8.14 (s, 1H), 8.02 (d, J = 5.2 Hz, 1H), 7.97 (s, 1H), 7.76 (d, J = 5.6 Hz, 1H), 7.71 (t, J = 8.0 Hz, 1H), 7.53 (d, J = 7.6 Hz, 1H), 7.44 (s, 1H), 7.30 (dd, J = 8.4, 6.8 Hz, 1H), 7.06 (dd, J = 11.6, 2.4 Hz, 1H), 6.98 (d, J = 7.6 Hz, 1H), 6.90–6.85 (m, 1H), 6.68 (d, J = 4.4 Hz, 1H), 5.61 (s, 2H), 5.15 (s, 1H), 3.94 (s, 2H), 3.76 (s, 3H), 1.30 (s, 6H).

[0316] Example 21: Preparation of compound Z-21

[0317]

[0318] Referring to the preparation method of Reference Example 19, except that 1-methyl-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyrazole is used to replace 1-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyrazole, compound Z-21 (9.63 mg) is obtained. MS m / z (ESI): 408.4, [M+1]; 1 H NMR(400MHz,DMSO-d 6 )δ9.64(s,1H),8.17(s,1H),8.01(d,J=5.2Hz,1H),7.79(d,J=5.2Hz,1H),7.53(d,J=2.0Hz,1H),7.39(s,1H),7.32–7.27(m,1H),7.05(dd,J=11.2,2.4Hz,1H),6.87(td,J=8.4,2.4Hz,1H),6.68(d,J=5.2Hz,1H),6.00(d,J=2.4Hz,1H),3.77(s,2H),3.75(s,3H),3.73(s,3H).

[0319] Example 22: Preparation of Compound Z-22

[0320]

[0321] Sodium hydride (59.70 mg, 2.49 mmol) was added to a solution of 1H-imidazo[4,5-b]pyridin-2-amine (185.39 mg, 1.38 mmol) and DMF (6 mL) at 0 °C. After the addition, the reaction was carried out at 0 °C for 30 minutes. Then compound 1-3 (200.00 mg, 552.83 μmol) was added to the above reaction solution, and the reaction was carried out at room temperature for 1.5 hours after the addition. The reaction solution was quenched with ice water, extracted with EA (10 mL × 2), washed with water (10 mL × 3), washed with saturated brine (10 mL), and purified by column chromatography (DCM: methanol = 100%:0% to 80%:20%). Separation by preparative liquid chromatography (preparative column: 21.2X250 mm C18 column; system: 10 mM NH4HCO3 H2O; wavelength: 254 / 214 nm) gave compound Z-22 (36.36 mg). MS m / z (ESI): 460.1 [M+1]; 1H NMR (400 MHz, DMSO-d6) δ 9.75 (s, 1H), 8.10 (d, J = 1.2 Hz, 1H), 8.07 (d, J = 5.2 Hz, 1H), 7.67 (d, J = 5.4 Hz, 1H), 7.52 (d, J = 6.0 Hz, 1H), 7.35 (s, 1H), 7.29 (dd, J = 8.4, 6.8 Hz, 1H), 7.23 (d, J = 7.6 Hz, 1H), 7.05 (dd, J = 11.4, 2.4 Hz, 1H), 6.87 (td, J = 8.4, 2.4 Hz, 1H), 6.77–6.72 (m, 1H), 6.59 (dd, J = 5.2, 1.2 Hz, 1H), 6.44 (s, 2H), 5.54 (s, 2H), 3.75 (s, 3H).

[0322] Example 23: Preparation of Compound Z-23

[0323]

[0324] Under nitrogen protection, sodium hydride (50.41 mg, 1.26 mmol, 60% purity) was added to a solution of 4-methyl-1H-1,2,3-triazole (87.28 mg, 1.05 mmol) and DMF (5 mL) at 0 °C. After the addition, the reaction was carried out at 0 °C for 15 minutes, and then a solution of compound 1-3 (380 mg, 1.05 mmol) and DMF (5 mL) was added. After the addition, the reaction was carried out at room temperature for 3 hours. The reaction solution was quenched with ice water and then extracted with EA (5 mL × 2). The combined organic phases were washed with water (10 mL × 3), saturated brine, dried over anhydrous sodium sulfate, and concentrated to obtain a crude product. The crude product was purified by column chromatography (PE:EA = 70%:30% - 50%:50%) to obtain compound Z-23 (84.35 mg). MS m / z (ESI): 409.1 [M+1]; 1H NMR (400 MHz, DMSO-d6) δ 9.78 (s, 1H), 8.16 (d, J = 1.2 Hz, 1H), 8.08 (d, J = 5.2 Hz, 1H), 7.72 (d, J = 5.4 Hz, 1H), 7.58 (s, 1H), 7.34 (s, 1H), 7.30 (dd, J = 8.2, 6.8 Hz, 1H), 7.06 (dd, J = 11.4, 2.4 Hz, 1H), 6.88 (td, J = 8.4, 2.4 Hz, 1H), 6.56 (dd, J = 5.2, 1.2 Hz, 1H), 5.52 (s, 2H), 3.76 (s, 3H), 2.22 (s, 3H).

[0325] Example 24: Preparation of Compound Z-24

[0326]

[0327] Step 1: 1,2-Difluoro-3-nitrobenzene (9.35 g, 58.77 mmol) and 1-hydroxycyclopropanecarboxylic acid (5 g, 48.98 mmol) were dissolved in DMF (50 mL), cooled to 0 °C, and 60% sodium hydride (4.90 g, 122.44 mmol) was added thereto. The reaction was stirred at room temperature for 0.5 hour and then heated to 80 °C for 3 hours. Water (80 mL) was added to quench the reaction, and the mixture was extracted with EA (60 mL). The aqueous phase was adjusted to pH 3 with 6N hydrochloric acid and then extracted with EA (80 mL × 3). The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain compound 24-1 (8.0 g). MS m / z (ESI): 242.0 [M+1]

[0328] Step 2: Dissolve compound 24-1 (8.0 g, 33.17 mmol) in ethanol (80 mL) and water (15 mL), then add reduced iron powder (9.26 g, 165.86 mmol) and ammonium chloride (8.87 g, 165.86 mmol) thereto. The reaction was stirred at 90 °C for 2 hours. Dilute with EA (100 mL), wash with saturated sodium chloride solution (100 mL×3), dry the organic phase with anhydrous sodium sulfate, filter, and concentrate under reduced pressure. Purify the obtained residue by silica gel column chromatography with eluent 0-30% EA in PE to obtain compound 24-2 (5.0 g). MS m / z (ESI): 194.1 [M+1]

[0329] Step 3: Dissolve compound 24-2 (2.7 g, 13.98 mmol) in THF (40 mL), then add borane (1 M, 41.93 mL) thereto. The reaction was stirred at 50 °C for 2 hours. Quench the reaction with methanol (15 mL), remove the solvent under reduced pressure, add 20 ml of 1N hydrochloric acid, heat to 50 °C and react for 10 minutes, cool to room temperature, add saturated sodium carbonate solution (50 mL), extract with EA, filter, and concentrate under pressure to obtain compound 24-3 (2.5 g). MS m / z (ESI): 180.1 [M+1].

[0330] Step 4: Dissolve compound 24-3 (2.5 g, 13.95 mmol) in acetonitrile (25 mL), then add NBS (4.97 g, 27.90 mmol) thereto. The reaction was stirred at room temperature for 2 hours. Dilute with EA (80 mL), wash with saturated sodium chloride solution (60 mL×3), dry the organic phase with anhydrous sodium sulfate, filter, and concentrate under reduced pressure. Purify the obtained residue by silica gel column chromatography with eluent 0-30% EA in PE to obtain compound 24-4 (2.0 mg). MS m / z (ESI): 337.8 [M+1].

[0331] Step 5: Dissolve compound 24-4 (1.8 g, 5.34 mmol) in THF (30 mL), cool to -78 °C, and add n-butyllithium (2.5 M, 4.70 mL) thereto. The reaction was stirred at -78 °C for 15 minutes, stop the reaction, quench with saturated sodium chloride solution (50 mL), extract with EA (60 mL), dry the organic phase with anhydrous sodium sulfate, filter, and concentrate under reduced pressure. Purify the obtained residue by silica gel column chromatography with eluent 0-30% EA in PE to obtain compound 24-5 (1.05 mg). MS m / z (ESI): 257.9 [M+1].

[0332] Step 6: Place compound 24-5 (550 mg, 2.13 mmol) and bis(pinacolato)diboron (1.08 g, 4.26 mmol) in a reaction flask, add DMSO (20 mL), and then add Pd(dppf)Cl 2 (155.93 mg, 213.11 μmol) and potassium acetate (627.43 mg, 6.39 mmol) thereto. The reaction was stirred at 120 °C for 2 h. Dilute with EA (60 mL), wash with saturated sodium chloride solution (60 mL x 3), dry the organic phase over anhydrous sodium sulfate, filter, and concentrate under reduced pressure to obtain compound 24-6 (500 mg). MS m / z (ESI): 306.1 [M+1]

[0333] Step 7: Dissolve compound 24-6 (500 mg, 1.64 mmol) and 2-chloro-5-fluoro-4-iodopyridine (463.99 mg, 1.80 mmol) in 1,4-dioxane (12 mL), and then add Pd(dppf)Cl 2 (59.95 mg, 81.93 μmol) and potassium carbonate (679.36 mg, 4.92 mmol) thereto. The reaction was stirred at 100 °C under microwave for 3 h. Dilute with EA (80 mL), wash with saturated sodium chloride solution (80 mL x 3), dry the organic phase over anhydrous sodium sulfate, filter, and concentrate under reduced pressure. Purify the obtained residue by silica gel column chromatography with eluent 0-50% EA in PE to obtain compound 24-7 (300 mg). MS m / z (ESI): 309.0 [M+1]

[0334] Step 8: Dissolve compound 24-7 (100 mg, 323.93 μmol) and (2-amino-4-pyridyl)methanol (60.32 mg, 485.89 μmol) in 1,4-dioxane (6 mL), and then add Pd 2 (dba) 3 (29.66 mg, 32.39 μmol), XantPhos (37.49 mg, 64.79 μmol) and cesium carbonate (211.08 mg, 647.86 μmol) thereto. The reaction was stirred at 100 °C for 3 h. Add EA (60 mL), wash with saturated sodium chloride solution (60 mL x 3), dry the organic phase over anhydrous sodium sulfate, filter, and concentrate under reduced pressure. Purify the obtained residue by silica gel column chromatography with eluent 0-80% EA in PE to obtain compound 24-8 (80 mg). MS m / z (ESI): 397.1 [M+1]

[0335] Step 9: Dissolve compound 24-8 (80 mg) (80 mg, 201.82 μmol) in DCM (15 mL), and then add thionyl chloride (120.05 mg, 1.01 mmol, 73.29 μL) thereto. The reaction was stirred at 40 °C for 3 hours. Add EA (80 mL), wash with saturated sodium chloride solution (60 mL x 3), dry the organic phase with anhydrous sodium sulfate, filter, and concentrate under reduced pressure to obtain compound 24-9 (80 mg). MS m / z (ESI): 415.0 [M+1].

[0336] Step 10: Dissolve compound 24-9 (80 mg, 192.85 μmol) and 1H-benzoimidazol-2-amine (38.52 mg, 289.27 μmol) in acetone (15 mL), and then add potassium carbonate (53.31 mg, 385.70 μmol) and potassium hydroxide (21.64 mg, 385.70 μmol) thereto. The reaction was stirred at 60 °C for 3 hours. Add EA (80 mL), wash with saturated sodium chloride solution (80 mL x 3), dry the organic phase with anhydrous sodium sulfate, filter, and concentrate under reduced pressure. The residue was purified by preparative liquid chromatography (preparative column: 21.2 X 250 mm C18 column; system: 10 mM NH 4 HCO 3 H 2 O; wavelength: 254 / 214 nm; gradient: 5%-95% acetonitrile change) to obtain the crude product, and obtain compound Z-24 (37.83 mg) MS m / z (ESI): 512.2 [M+1]; 1 1H NMR (400 MHz, DMSO-d6) δ 9.73 (s, 1H), 8.08 (d, J = 1.2 Hz, 1H), 8.06 (d, J = 5.2 Hz, 1H), 7.68 (d, J = 5.6 Hz, 1H), 7.42 (s, 1H), 7.13 (d, J = 7.6 Hz, 1H), 6.98 (d, J = 7.6 Hz, 1H), 6.91 (td, J = 7.6, 1.2 Hz, 1H), 6.80 (td, J = 7.6, 1.2 Hz, 1H), 6.60 (dd, J = 8.4, 5.6 Hz, 1H), 6.54–6.43 (m, 4H), 5.63 (s, 1H), 5.23 (s, 2H), 3.19 (d, J = 2.8 Hz, 2H), 0.93 (t, J = 6.0 Hz, 2H), 0.69 (t, J = 6.0 Hz, 2H).

[0337] Example 25: Preparation of compound Z-25

[0338]

[0339] Compound 24-9 (70 mg, 168.74 μmol) and 1-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyrazole (105.33 mg, 506.23 μmol) were dissolved in 1,4-dioxane (8 mL) and water (2 mL), and then Pd(PPh 3 ) 4 (29.25 mg, 25.31 μmol) and cesium carbonate (164.94 mg, 506.23 μmol) were added thereto. The reaction was stirred at 80 °C for 6 hours. EA (30 mL) was added, and the mixture was washed with saturated sodium chloride solution (30 mL x 3). The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by preparative liquid chromatography (preparative column: 21.2 X 250 mm C18 column; system: 10 mM NH4HCO3 H2O; wavelength: 254 / 214 nm; gradient: 5%-95% acetonitrile change) to obtain compound Z-25 (8.70 mg). MS m / z (ESI): 461.1 [M+1]; 1 1H NMR (400 MHz, DMSO-d6) δ 9.72 (s, 1H), 8.17 (s, 1H), 8.07 (d, J = 5.2 Hz, 1H), 7.76 (d, J = 5.6 Hz, 1H), 7.41 (s, 1H), 7.29 (d, J = 1.6 Hz, 1H), 6.72–6.55 (m, 2H), 6.54–6.42 (m, 1H), 5.99 (d, J = 1.6 Hz, 1H), 5.66 (s, 1H), 3.98 (s, 2H), 3.67 (s, 3H), 3.20 (s, 2H), 0.94 (t, J = 6.0 Hz, 2H), 0.70 (t, J = 6.0 Hz, 2H).

[0340] Example 26: Preparation of Compound Z-26

[0341]

[0342] Referring to the preparation method in Step 10 of Reference Example 24, the difference is that N-methyl-1H-benzoimidazol-2-amine was used instead of 1H-benzoimidazol-2-amine to obtain compound Z-26. MS m / z (ESI): 526.3 [M+1]; 11H NMR (400 MHz, DMSO-d6) δ 9.74 (s, 1H), 8.09 (d, J = 1.2 Hz, 1H), 8.07 (d, J = 5.2 Hz, 1H), 7.70 (d, J = 5.6 Hz, 1H), 7.40 (s, 1H), 7.22 (d, J = 8.0 Hz, 1H), 7.02 (d, J = 7.2 Hz, 1H), 6.94 (td, J = 7.6, 1.2 Hz, 1H), 6.84 (td, J = 7.6, 1.2 Hz, 1H), 6.76 (d, J = 4.8 Hz, 1H), 6.61 (dd, J = 8.8, 5.6 Hz, 1H), 6.57 - 6.46 (m, 2H), 5.64 (s, 1H), 5.22 (s, 2H), 3.21 (d, J = 2.0 Hz, 2H), 2.92 (d, J = 4.4 Hz, 3H), 0.95 (t, J = 6.4 Hz, 2H), 0.71 (t, J = 6.4 Hz, 2H).

[0343] Example 27: Preparation of Compound Z-27

[0344]

[0345] Step 1: Referring to the preparation method of Step 8 in Example 24, except that diphenylmethanimine is used to replace (2-amino-4-pyridyl)methanol, to obtain Compound 27-1. MS m / z (ESI): 454.1 [M+1]

[0346] Step 2: Dissolve Compound 27-1 (700 mg, 1.54 mmol) in methanol (30 mL), and then add hydroxylamine hydrochloride (536.3 mg, 7.72 mmol) thereto. The reaction is stirred at room temperature for 2 hours. The solvent is removed under reduced pressure, ethyl acetate (50 mL) is added, and it is washed with saturated sodium bicarbonate solution and saturated sodium chloride solution respectively. The organic phase is dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue is separated and purified by silica gel column chromatography to obtain Compound 27-2 (350 mg). MS m / z (ESI): 290.1 [M+1].

[0347] Step 3: Place compound 27-2 (100 mg, 345.69 μmol) and compound 4-8 (171.22 mg, 622.24 μmol) in a microwave tube. Add toluene (6 mL), N-methylpyrrolidone (0.6 mL), and then add chloro(2-dicyclohexylphosphino-2',4',6'-triisopropyl-1,1'-biphenyl)[2-(2-aminoethylphenyl)]palladium(II) (51.08 mg, 69.14 μmol), X-PHOS (32.96 mg, 69.14 μmol), and potassium phosphate (366.89 mg, 1.73 mmol). The reaction is stirred in a microwave reactor at 120 °C for 6 hours. Add ethyl acetate (50 mL), wash the organic phase with saturated brine (50 mL × 3), dry over anhydrous sodium sulfate, filter, and concentrate under reduced pressure. The residue is purified by preparative liquid chromatography (preparative column: 21.2 × 250 mm C18 column; system: 10 mM NH 4 HCO 3 H 2 O; wavelength: 254 / 214 nm; gradient: 5% - 95% acetonitrile change) to obtain compound Z-27 (32.85 mg). MS m / z (ESI): 484.1 [M+1]; 1 H NMR (400 MHz, DMSO-d 6 ) δ 9.80 (s, 1H), 8.23 (d, J = 1.2 Hz, 1H), 8.16 (d, J = 5.2 Hz, 1H), 7.77 (d, J = 5.2 Hz, 1H), 7.65 (s, 1H), 6.93 (dd, J = 5.2, 1.2 Hz, 1H), 6.65 (dd, J = 8.8, 5.6 Hz, 1H), 6.51 (dd, J = 10.4, 8.4 Hz, 1H), 5.68 (s, 1H), 4.53 (dd, J = 42.0, 13.6 Hz, 2H), 3.65–3.53 (m, 1H), 3.44 - 3.36 (m, 1H), 3.22 (d, J = 2.8 Hz, 2H), 3.19–3.12 (m, 1H), 3.04–2.92 (m, 1H), 2.06 - 1.96 (m, 1H), 1.92 - 1.82 (m, 1H), 0.96 (t, J = 6.0 Hz, 2H), 0.72 (t, J = 6.0 Hz, 2H).

[0348] Example 28: Preparation of Compound Z-28 6618

[0349]

[0350] Step 1-2: Refer to the preparation method of Step 1-2 in Example 3, with the difference that 2-chloro-4-(chloromethyl)pyridine is used to replace Compound 1-3, obtaining Compound 28-1. MS m / z (ESI): 260.1 [M+1].

[0351] Step 3-4: Refer to the preparation method of Step 5-6 in Example 1, with the difference that Compound 28-1 is used to replace Compound 1-4, obtaining Compound 28-4. MS m / z (ESI): 391.1 [M+1].

[0352] Step 5: Using Compound 28-4 and Compound 27-2 as raw materials, refer to the preparation method of Step 3 in Example 27, obtaining Compound 28-5, MS m / z (ESI): 644.3 [M+1].

[0353] Step 6: Refer to the preparation method of Step 7 in Example 1, obtaining Compound 28-6. MS m / z (ESI): 544.2 [M+1].

[0354] Step 7: Dissolve Compound 28-6 (160 mg, 294.34 μmol) in methanol (12 mL) and water (3 mL), then add sodium hydroxide (58.8 mg, 1.47 mmol) thereto. After the reaction is completed, adjust to pH = 3-4 with 2N dilute hydrochloric acid, evaporate to dryness under reduced pressure, dry, obtaining Compound 28-7 (150 mg). MS m / z (ESI): 530.2 [M+1].

[0355] Step 8: Dissolve Compound 28-7 (150.00 mg, 141.63 μmol) in acetonitrile (10 mL), then add N,N,N',N'-tetramethylchloroformamidinium hexafluorophosphate (79.48 mg, 283.26 μmol) and N-methylimidazole (34.88 mg, 424.88 μmol) thereto. The reaction is stirred at room temperature for 2 hours. Dilute with ethyl acetate (50 mL), wash the organic phase with saturated sodium chloride solution (30 mL x 3), dry over anhydrous sodium sulfate, filter, and concentrate under reduced pressure. The residue is purified by preparative liquid chromatography (preparative column: 21.2X250 mm C18 column; system: 10 mM NH4HCO3 H2O; wavelength: 254 / 214 nm; gradient: 5%-95% acetonitrile change), obtaining Compound Z-28 (30.84 mg). MS m / z (ESI): 512.2 [M+1]; 1 HNMR(400MHz,DMSO-d 6)δ9.89(s,1H),8.24 - 8.19(m,2H),7.80–7.65(m,2H),6.92(dd,J=5.2,1.2Hz,1H),6.65(dd,J=8.4,5.6Hz,1H),6.51(dd,J=10.4,8.8Hz,1H),5.68(s,1H),4.79(dd,J=28.8,13.6Hz,2H),3.69–3.57(m,1H),3.26–3.11(m,3H),2.38–2.18(m,2H),2.16–1.99(m,2H),0.96(t,J=6.0Hz,2H),0.72(t,J=6.0Hz,2H).

[0356] Test Example 1: Activity Inhibition Test of CDK Family Kinases

[0357] In the following LANCE Ultra test method, the kinase reagent was purchased from Carna Bioscience, the reaction substrate and detection reagent were purchased from PerkinElmer, and the remaining reagents were purchased from Thermo scientific.

[0358] The inhibitory effect of the test substance on the kinase activities of CDK1 / CycB (Carna bioscience, #04 - 102), CDK2 / CycA (Carna bioscience, #04 - 103), and CDK9 / CycT (Carna bioscience, #04 - 110) was determined using the LANCE Ultra method.

[0359] The kinase activity test used a 10 μL system, which included: a CDK kinase dilution solution, a substrate dilution solution mixed with Ulight-Myelicbasic protein (PerkinElmer, #TRF - 0109, hereinafter referred to as U - MBP) and ATP (Thermo scientific, #PV3227), and the compound prepared in the above example of the present invention (i.e., the test substance). Each kinase in the test included three test groups: a background group (Blank), a non - inhibition group (PC), and a compound test group (Test). The components included in each test group were as follows:

[0360] Kinase Substrate Compound Blank 1.33x Reaction Buffer Substrate Diluent Only 2% DMSO PC Kinase Diluent Substrate Diluent Only 2% DMSO Test Kinase Diluent Substrate Diluent Compound Diluent

[0361] The working concentrations of the components in the Test group for different kinase reactions were as follows:

[0362] Compound: Dissolve the compound to be tested at 10 mM at room temperature and perform gradient dilution with DMSO, and then dilute it with deionized water to a 4x compound working solution with the DMSO content being 2%. The highest concentration of the compound used in the CDK1 and CDK2 tests is 10 μM, and for CDK9 it is 1 μM.

[0363] 1.33x Reaction Buffer: Composed of 26.7 mM MOPS, 6.67 mM MgCl 2 and 0.0133% Tween-20. After preparation, store it in the refrigerator at 4 °C in the dark. Add freshly prepared DTT to a final concentration of 5.33 mM before use.

[0364]

[0365] The working concentration of DMSO in the reaction is 0.5%.

[0366] After mixing the above components, place them on a shaker and incubate in the dark at room temperature for 1 hour. Then add 10 μL of the detection solution to all test groups (including Blank, PC, and Test groups).

[0367] The 10 μL detection solution contains the following components: 16 mM EDTA (Thermo scientific, #15575), 1 nM phosphorylated U-MBP protein antibody (Perkin Elmer, #TRF-0201), and 1x detection buffer (Perkin Elmer, #CR97-100).

[0368] After adding the detection solution, place it on a shaker and continue to incubate in the dark at room temperature for 1 hour. After the incubation, use the Victor X5 fluorescence microplate reader from PerkinElmer to read the signal. The excitation wavelength is 320 nm, and the emission wavelengths are 615 nm and 665 nm. The method for calculating the inhibition rate is as follows:

[0369] 1. Calculate the value of 665 nm / 615 nm (hereinafter referred to as the Ratio value) for all groups, and calculate the inhibition rate based on the Ratio value of each group;

[0370] 2. Inhibition rate = (PC Ratio – Test Ratio ) / (PC Ratio – Blank Ratio ) * 100%;

[0371] 3. Use XLFIT 5.0 software (IDBS, UK) for fitting. Use the logarithm of the compound concentration as the X-axis and the inhibition rate as the Y-axis, and use the four-parameter model to calculate the half-maximal inhibitory concentration IC 50 .

[0372] Table 1 Inhibitory activities of compounds against CDK1, CDK2 and CDK9

[0373]

[0374] As can be seen from Table 1, the compounds of the embodiments of the present invention have relatively high selective inhibitory activity against CDK9.

[0375] Test Example 2: Cell proliferation assay

[0376] MV-4-11 cells (20,000 cells / well) were seeded in a 96-well plate (Corning #3916) and cultured. The culture medium was 90 μL / well (IMDM containing 10% fetal bovine serum (FBS), Gibco #10099-141C). After culturing for 16 h, 10 μL of the compounds of the present invention prepared at different concentrations (final concentration of DMSO was 0.5% (v / v)) was added. After continued culturing for 3 days, 50 μL of CellTiter-Glo (Promega #G7558) was added to each well, and the plate was placed on a shaker at 80 rpm, 25 °C for 15 min. The fluorescence value was detected using a Tecan Infinite pro3000. The test results are shown in the table.

[0377] Table 2: IC 50 value of the inhibitory activity of the compounds against the proliferation of MV-4-11 cells

[0378]

[0379]

[0380] As can be seen from Table 2, the compounds of the embodiments of the present invention have good inhibitory activity against the proliferation of MV-4-11 cells.

[0381] Test Example 3: Pharmacokinetic study in mice

[0382] The LC / MS / MS method was used to determine the drug concentration in plasma at different time points after intragastric administration of the compounds of the present invention to mice, study the pharmacokinetic behavior of the compounds of the present invention in mice, and evaluate their pharmacokinetic characteristics.

[0383] Experimental protocol:

[0384] Test animals: Healthy adult male ICR mice (weight 30 - 40 g, 12 mice in the intragastric administration group were fasted overnight and allowed free access to water and food 4 h after administration), provided by Beijing Vital River Laboratory Animal Co., LTD;

[0385] Route of administration and dosage: ICR mice were administered by tail vein gavage (10 mg / kg, 5% DMSO, pH 4.5 20% Captisol).

[0386] Blood sample collection: Animals meeting the experimental requirements were selected and weighed and marked before administration. Before collecting blood samples, the mice were restrained. For each mouse administered, at the predetermined blood collection time points (gavage administration: blood was collected at 0.083, 0.25, 0.5, 1, 2, 4, 6, 7.5, 24 h after administration, a total of 9 time points), approximately 100 μL of blood was collected by orbital bleeding. The blood was transferred to a 1.5 mL tube pre-added with K 2 EDTA, centrifuged for 4 min (8000 rpm, 4 °C), and the plasma was taken out. The whole process was completed within 15 min after blood collection. All samples were stored in a -20 °C refrigerator until sample analysis. The drug concentration was determined by LC / MS / MS method. For the compounds in some embodiments of the present invention, the pharmacokinetic property parameters in mice under the same dose and route of administration are shown in Table 3 as follows:

[0387] Table 3 Pharmacokinetic parameters of the compound in mice

[0388] Pharmacokinetic Parameter Z-1 Z-4 Z-17 <![CDATA[Oral maximum plasma drug concentration C max (ng / mL)]]> 773 2763 1723 <![CDATA[Oral area under the curve AUC 0-t (hr*ng / mL)]]> 879 2726 757

[0389] Test Example 4: In vivo efficacy experiment

[0390] The in vivo efficacy experiment was carried out on BALB / c nude mice subcutaneously implanted with a xenograft (CDX) of a human tumor cell line derived from MV4-11 acute myeloid leukemia patients.

[0391] Experimental protocol: BALB / c nude mice, female, 6 - 10 weeks old, weighing about 20 - 23 g, were kept in a special pathogen-free environment and in single ventilated cages (4 mice per cage, 2 cages per group, 8 mice in total). All cages, bedding, and water were disinfected before use. All animals had free access to standard certified commercial laboratory diets. A total of 85 mice purchased from the Experimental Animal Business Department of Shanghai Institute of Planned Parenthood Research (No. 3577 Jinke Road, Pudong, Shanghai) were used for the study. Each mouse was subcutaneously implanted with tumor cells (1×10 7 0.1 ml + Matrigel 0.1 ml) for tumor growth. When the average tumor volume reached approximately 165 cubic millimeters, administration began. The test compound was administered by oral gavage daily at a dose of 10 mpk. The anti-tumor efficacy was determined by dividing the average tumor increased volume of the animals treated with the compound by the average tumor increased volume of the untreated animals.

[0392] The tumor volume was measured twice a week with a two-dimensional caliper, and the volume was measured in cubic millimeters. Tumor volume TV = 0.5a × b 2Among them, a is the long diameter of the tumor, and b is the short diameter of the tumor.

[0393] Relative tumor growth rate T / C(%), that is, at a certain time point, the percentage value of the relative tumor volume of the treatment group and the control group. The calculation formula is as follows: T / C% = T RTV / C RTV ×100% (T RTV : average RTV of the treatment group; C RTV : average RTV of the vehicle control group; RTV = V t / V 0 ,V 0 is the tumor volume of the animal at the time of grouping, and Vt is the tumor volume of the animal after treatment).

[0394] The weight change (%) of the tumor-bearing animals is calculated as follows: (weight at measurement - weight at grouping) / weight at grouping × 100.

[0395] The effect of compound Z-1 on the tumor volume of animals is specifically as Figure 1 shown, and the relative tumor growth rate is specifically as Figure 2 shown. Figure 1 and Figure 2 The data in show that compound Z-1 exhibits a significant effect of inhibiting tumor growth and has anti-tumor efficacy.

[0396] The effect of compound Z-1 on the weight change of tumor-bearing animals is specifically as Figure 3 shown. The weight of the tumor-bearing animals did not decrease significantly before and after drug treatment, and no animals died.

[0397] All the documents mentioned in the present invention are cited in this application as references, just as if each document was cited separately as a reference. In addition, it should be understood that after reading the above teachings of the present invention, those skilled in the art can make various changes or modifications to the present invention, and these equivalent forms also fall within the scope defined by the appended claims of this application.

Claims

1. A compound of formula (I), or a pharmaceutically acceptable salt thereof: (I)； In the formula, (R 01 ) n indicates that the hydrogen on the pyridine ring is substituted by n R 01 , where n is 0, 1, 2 or 3; each R 01 is the same or different and is independently a halogen; (R 02 ) m represents that the hydrogen on the pyridine ring is substituted by m R 02 groups, where m is 0; Q is a 5- to 10-membered heteroaryl group, and the 5- to 10-membered heteroaryl group is unsubstituted or substituted by 1, 2 or 3 substituents each independently selected from the group consisting of: deuterium, halogen, cyano, hydroxy, carboxy, C 1-3 alkyl, C 1-3 alkoxy, halo-C 1-3 alkyl, halo-C 1-3 alkoxy, NR a0 R b0 , -C(O)NR a0 R b0 , substituted or unsubstituted -C 1-4 alkyl-5- to 6-membered heteroaryl; wherein substitution means that 1, 2 or 3 hydrogens in the group are independently substituted by substituents selected from the group consisting of: halogen, cyano, hydroxy, carboxy, C 1-3 alkyl, C 1-3 alkoxy, hydroxy-substituted C 1-3 alkyl, NR a0 R b0 ; Or Q is a structure represented by formula (a), formula (b) or formula (c): 、 、 ； wherein Z 1 is CH 2 or C(O); Z 2 is (CR c R d ) t0 ; each R c is the same or different, each R d is the same or different, and R c and R d are each independently hydrogen, cyano, hydroxy, carboxy, halogen, C 1-8 alkyl, or C 1-8 alkoxy; t0 is 1, 2, 3, 4 or 5; R 1a and R 2a are each independently hydrogen or C 1-3 alkyl; or R 1a is connected to R 2a to form a 5- to 8-membered saturated monocyclic heterocycle; the 5- to 8-membered saturated monocyclic heterocycle is unsubstituted or substituted with 1, 2, 3, or 4 R s1 ; where each R s1 is the same or different and is independently selected from: halogen, cyano, hydroxy, C 1-3 alkyl, C 1-3 alkoxy, halo-C 1-3 alkyl, halo-C 1-3 alkoxy; R 3a and R 4a each independently represents hydrogen, cyano, C 1-8 alkyl; or R 3a and R 4a are linked to form a 5- to 8-membered heteroalkenyl ring or a 5- to 6-membered heteroaryl ring; the 5- to 8-membered heteroalkenyl ring and the 5- to 6-membered heteroaryl ring are unsubstituted or substituted by 1, 2, 3 or 4 R s2 ; where each R s2 is the same or different and is independently selected from: halogen, cyano, hydroxy, C 1-3 alkyl, C 1-3 alkoxy, halo-C 1-3 alkyl, halo-C 1-3 alkoxy; R 5a is hydrogen; R 6a Connect with R 7a to form a 5- to 8-membered heteroalkenyl ring; the 5- to 8-membered heteroalkenyl ring is unsubstituted or substituted by two R s3 substituents; any two R s3 on the same ring atom are connected to form a 3- to 7-membered saturated monocyclic ring; the 3- to 7-membered saturated monocyclic ring is unsubstituted; L is CH 2 ; R 1 、R 2 、R 3 Each independently is hydrogen, halogen or C 1-8 alkoxy group; R 4 、R 5 are each independently hydrogen; or R 4 is connected to R 5 to form a fused 3- to 7-membered partially unsaturated monocyclic heterocycle; the fused 3- to 7-membered partially unsaturated monocyclic heterocycle is unsubstituted or substituted by 1, 2, 3, or 4 R s4 ; wherein each R s4 is the same or different, and any two R s4 on the same ring atom are connected to form a 3- to 6-membered saturated monocyclic ring; the 3- to 6-membered saturated monocyclic ring is unsubstituted; R a0 、R b0 are each independently hydrogen, C 1-3 alkyl or acetyl group.

2. The compound according to claim 1, or a pharmaceutically acceptable salt thereof, wherein, n is 1; R 01 is fluorine.

3. The compound according to claim 1, or a pharmaceutically acceptable salt thereof, wherein, For 。 4. The compound according to claim 1, or a pharmaceutically acceptable salt thereof, wherein, When Q is a 5- to 10-membered heteroaryl, the 5- to 10-membered heteroaryl is a 5- to 6-membered heteroaryl or an 8- to 10-membered heteroaryl.

5. The compound according to claim 1, or a pharmaceutically acceptable salt thereof, wherein, The 5- to 6-membered heteroaryl is selected from: imidazolyl, oxazolyl, pyrrolyl, pyrazolyl, triazolyl; the 8- to 10-membered heteroaryl is selected from: benzimidazolyl, purinyl.

6. The compound according to claim 1, or a pharmaceutically acceptable salt thereof, wherein, In formula (a), Z 2 is (CH 2 ) t0 ; t 0 is 1, 2, 3 or 4.

7. The compound according to claim 1, or a pharmaceutically acceptable salt thereof, wherein, Formula (a) is selected from any of the following structures: 、 、 、 、 、 。 8. The compound according to claim 1, or a pharmaceutically acceptable salt thereof, wherein, The 5- to 8-membered saturated monocyclic heterocycle is imidazolidine.

9. The compound according to claim 1, or a pharmaceutically acceptable salt thereof, wherein, The R 3a connected to R 4a forms a 5- to 8-membered heteroalkenyl ring which is a 4,5-dihydro-1H-imidazole ring.

10. The compound according to claim 1, or a pharmaceutically acceptable salt thereof, wherein, The R 6a is connected to R 7a to form a 5- to 8-membered heteroalkenyl ring which is a 5,6-dihydro-4H-1,3-oxazine ring.

11. The compound according to claim 1, or a pharmaceutically acceptable salt thereof, wherein, Q is selected from any of the following structures: 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 。 12. The compound according to claim 1, or a pharmaceutically acceptable salt thereof, wherein, R 1 is hydrogen or methoxy group.

13. The compound according to claim 1, or a pharmaceutically acceptable salt thereof, wherein, R 3 is fluorine.

14. The compound according to claim 1, or a pharmaceutically acceptable salt thereof, wherein, R 4 、R 5 Each independently represents hydrogen.

15. The compound according to claim 1, or a pharmaceutically acceptable salt thereof, wherein, Structure Selected from any of the following structures: 、 。 16. The compound according to claim 1, or a pharmaceutically acceptable salt thereof, wherein, The compound of formula (I) is any one of the following compounds: 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 。 17. A pharmaceutical composition, wherein, It comprises the compound according to any one of claims 1-16, or a pharmaceutically acceptable salt thereof; and a pharmaceutically acceptable carrier.

18. Use of the compound according to any one of claims 1-16, or a pharmaceutically acceptable salt thereof or the pharmaceutical composition according to claim 17 in the preparation of a drug for treating or preventing a disease related to or mediated by CDK9 activity.

19. The use according to claim 18, wherein, The disease related to or mediated by CDK9 activity is a hyperproliferative disease.

20. The use according to claim 19, wherein, The disease related to or mediated by CDK9 activity is cancer.

21. The use according to claim 20, wherein, The cancer is selected from pancreatic cancer, breast cancer, ovarian cancer, cervical cancer, and leukemia.

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